Michihiro Hayashi

Background. Although living donor liver transplantation for small pediatric patients is increasingly accepted, its expansion to older/larger patients is still in question because of the lack of sufficient information on the impact of graft size mismatching. Methods. A total of 276 cases of living donor liver transplantation, excluding ABO-incompatible, auxiliary, or secondary transplants, were reviewed from graft size matching. Forty-three cases were highly urgent cases receiving intensive care preoperatively. Cases were categorized into five groups by graft-to-recipient weight ratio (GRWR): extra-small-for-size (XS; GRWR<0.8%, 17 elective and 4 urgent cases), small (S; 0.8≤GRWR<1.0%, 21 and 7), medium (M; 1.0≤GRWR<3.0%, 119 and 19), large (L; 3.0≤GRWR<5.0%, 67 and 10), and extra-large (XL; GRWR≥5.0%, 9 and 3). Results. Smaller-for-size grafts were associated not only with larger and older recipients, but also with rather older donors. Posttransplant bilirubin clearance was delayed and aspartate aminotransferase corected by relative graft size was higher in XS and S. Posttransplant hemorrhage and intestinal perforation were more frequent in XS and S, and vascular complications and acute rejection were more frequent in larger-for-size grafts. Consequently, graft survival in XS (cumulative 58% and actuarial 42% at 1 year) and S (76% and 74%) was significantly lower compared with that in M (93% and 92%) in elective cases. Graft survival in L (83% and 82%) and XL (75% and 71%) did not reach statistical significance. Conclusions. The use of small-for-size grafts (less than 1% of recipient body weight) leads to lower graft survival, probably through enhanced parenchymal cell injury and reduced metabolic and synthetic capacity. Although large-for-size grafts are associated with some anatomical and immunological disadvantages, the negative impact is less pronounced.

For the sake of donor safety in living donor liver transplantation (LDLT), the left lobe is currently being used most often for the graft. However, size mismatch has been a major obstacle for an expansion of the indication for LDLT to larger-size recipients, because a left lobe graft is not safe enough for them.In 1998, LDLT using a right lobe graft was introduced and performed on 26 recipients to overcome the small-for-size problem. The right lobe, which does not include the middle hepatic vein of the donor, was used. Initially, indication for right lobe LDLT was basically defined as an estimated left lobe graft volume/recipient body weight ratio (GRWR) of <0.8%, which was later raised to <1.0%.All the donors recovered from the operation without persistent complications. Two donors with transient bile leakage were successfully treated with a conservative approach. A right lobectomy resulted in more blood loss (337+/-175 ml), and a longer operative time (6.67+/-0.85 hr) than a lateral segmentectomy, but not a left lobectomy. Grafts with a GRWR >0.8% were implanted in all recipients, except for two, who received relatively smaller right lobes (GRWR of 0.68% and 0.66%). In one of these two, the right lobe from the donor was used as the orthotopic auxiliary graft. Postoperative transitory increases in total bilirubin and aspartate transaminoferase for right lobe donors were higher than those for the left lateral segmentectomy. Nineteen recipients (73.1%) were successfully treated with this procedure. The causes of death were not specific for right lobe LDLT, except for one patient with a graft that had multiple hepatic venous orifices. These multiple and separate anastomoses of the hepatic veins caused an outflow block as a result of a positional shift of the graft, which finally led to graft loss.Our experience suggests that right lobe grafting is a safe and effective procedure, resulting in the expansion of the indication for LDLT to large-size recipients. How to deal with the possible variation in the anatomy of the right lobe graft should be given attention throughout the procedure.

YAMAOKA, YOSHIO; WASHIDA, MASANOBU; HONDA, KAZUO; TANAKA, KOICHI; MORI, KEIICHIRO; SHIMAHARA, YASUYUKI; OKAMOTO, SHINGO; UEDA, MIKIKO; HAYASHI, MICHIHIRO; TANAKA, AKIRA; MORIMOTO, TAISUKE; OZAWA, KAZUE Author Information

Some reported studies have indicated the possibility of immunosuppression withdrawal in cadaveric liver transplantation. The aim of this study was to evaluate the possibility and feasibility of weaning living donor liver transplant recipients from immunosuppression.From June of 1990 to October of 1999, 63 patients were considered to be weaned from immunosuppression. They consisted of 26 electively weaned patients and 37 either forcibly or incidentally weaned patients (nonelective weaning) due to various causes but mainly due to infection. Regarding elective weaning, we gradually reduced the frequency of tacrolimus administration for patients who survived more than 2 years after transplantation, maintained a good graft function, and had no rejection episodes in the preceding 12 months. The frequency of administration was reduced from the conventional b.i.d. until the start of weaning to q.d., 4 times a week, 3 times a week, twice a week, once a week, twice a month, once a month, and finally, the patients were completely weaned off with each weaning period lasting from 3 to 6 months. The reduction method of nonelective weaning depended on the clinical course of each individual case. When the patients were clinically diagnosed to develop rejection during weaning, then such patients were treated by a reintroduction of tacrolimus or an additional steroid bolus when indicated.Twenty-four patients (38.1%) achieved a complete withdrawal of tacrolimus with a median drug-free period of 23.5 months (range, 3-69 months). Twenty-three patients (36.5%) are still being weaned at various stages. Sixteen patients (25.4%) encountered rejection while weaning at median period of 9.5 months (range, 1-63 months) from the start of weaning. All 16 were easily treated with the reintroduction of tacrolimus or additional steroid bolus therapy.We were able to achieve a complete withdrawal of immunosuppression in some selected patients. Although the mechanism of graft acceptance in these patients has yet to be elucidated, we believe that a majority of long-term patients undergoing living donor liver transplantation may, thus, be potential candidates to be successfully weaned from immunosuppression.

YAMAOKA, YOSHIO; WASHIDA, MASANOBU; HONDA, KAZUO; TANAKA, KOICHI; MORI, KEIICHIRO; SHIMAHARA, YASUYUKI; OKAMOTO, SHINGO; UEDA, MIKIKO; HAYASHI, MICHIHIRO; TANAKA, AKIRA; MORIMOTO, TAISUKE; OZAWA, KAZUE Author Information

Background. Hepatic steatosis affects graft function as well as postoperative recovery of donors in living donor liver transplantation. Liver macrovesicular steatosis in living donors was assessed using quantitative X-ray computed tomography (CT) analysis and histological examination of intraoperative liver biopsy. Methods. A total of 266 living donors with complete pretransplant CT data and intraoperative “time 0” biopsy were included in the study. Liver biopsy specimen obtained during donor operation was examined for macrovesicular steatosis and was classified as none; mild (<30%); moderate (30%–60%); or severe (>60%). Liver-to-spleen CT attenuation values ratio (L/S ratio) on noncontrast-CT was evaluated for its usefulness as an index of hepatic steatosis in comparison with other parameters including body mass index (BMI) and serum liver function tests (gamma-glutamyl transpeptidase, alanine aminotransferase, aspartate aminotransferase, cholinesterase, and total cholesterol) using receiver operating characteristic (ROC) analysis. Results. Histological grade of macrovesicular steatosis was none in 198 patients (74.4%), mild in 50 (18.8%), moderate in 15 (5.7%), and severe in 3 (1.1%). The median L/S ratios for the respective histological grades were 1.20 (range: 1.00–1.46), 1.12 (0.83–1.37), 1.01 (0.74–1.21), and 0.90 (0.70–0.99) (P<0.0001). The ROC curve for L/S ratio was located closest to the upper left corner, and the area under the curve of L/S ratio was significantly larger than that of any other preoperative variables. Conclusion. L/S ratio calculated from preoperative CT can be a useful tool to discriminate hepatic macrovesicular steatosis. Based on the present results, the optimal cut-off value for L/S ratio to exclude more than moderate steatosis would be 1.1.

In the central nervous system (CNS), the expression of molecules is strictly regulated during development. Control of the spatiotemporal expression of molecules is a mechanism not only to construct the functional neuronal network but also to adjust the network in response to new information from outside of the individual, i.e., through learning and memory. Among the functional molecules in the CNS, one of the best-studied groups is the neurotrophins, which are nerve growth factor (NGF)-related gene family molecules. Neurotrophins include NGF, brain-derived neurotrophic factor (BDNF), neurotrophin 3 (NT-3), and NT-4/5 in the mammal. Among neurotrophins and their receptors, BDNF and tropomyosin-related kinases B (TrkB) are enriched in the CNS. In the CNS, the BDNF-TrkB signaling pathway fulfills a wide variety of functions throughout life, such as cell survival, migration, outgrowth of axons and dendrites, synaptogenesis, synaptic transmission, and remodeling of synapses. Although the same ligand and receptor, BDNF and TrkB, act in these various developmental events, we do not yet understand what kind of mechanism provokes the functional multiplicity of the BDNF-TrkB signaling pathway. In this review, we discuss the mechanism that elicits the variety of functions performed by the BDNF-TrkB signaling pathway in the CNS as a tool of pharmacological therapy.

Aim Microvascular invasion ( MVI ) is an important risk factor for early recurrence of hepatocellular carcinoma ( HCC ), but preoperative prediction of MVI is difficult. Methods A retrospective review was undertaken on 167 patients with primary solitary HCC who underwent initial hepatectomy. Independent predictors of MVI were identified, and factors affecting disease‐free survival in patients with MVI were clarified. Results Of the 167 patients, 20 patients (12%) had MVI . Recurrence rates of HCC after hepatectomy in MVI patients were significantly worse than in patients without MVI ( P &lt; 0.0361). Univariate analysis revealed that positive L3‐ AFP , PIVKA‐II ≥ 150 mAU/mL and tumor size ≥3 cm preoperatively were associated with positive MVI . On multivariate analysis, independent predictors of MVI were PIVKA‐II ≥ 150 mAU/mL (odds ratio [ OR] , 5.19; 95% confidence interval [95% CI ], 1.44–24.87; P = 0.0109) and positive L3 ‐ AFP ( OR , 3.47; 95% CI , 1.19–10.75; P = 0.0229). Among the MVI ‐positive group, the 1‐, 2‐ and 3‐year disease‐free survival rates were 78%, 58%, and 58% in patients with surgical margin ( SM ) ≥ 10 mm and 38%, 29%, and 29% in those with SM &lt; 10 mm, respectively ( P = 0.0263). Conclusions Patients with PIVKA‐II ≥ 150 mAU/mL and positive L3 ‐ AFP on preoperative examination are at high risk for MVI .

Hepatocellular carcinoma (HCC) often recurs after curative hepatectomy; and early recurrence after hepatectomy (ERAH) is associated with poor prognosis. This study aimed to clarify risk factors and disease patterns for ERAH. We retrospectively analyzed clinicopathological factors of 232 patients who underwent initial curative hepatectomies for HCC between April 2000 and March 2013, and examined associated risk factors and early recurrence patterns by liver function status (as indicated by indocyanine green retention rate at 15 min [ICGR15]). Patients who experienced recurrence within 6 months after hepatectomy (i.e., ERAH) had significantly shorter survival than those with longer disease-free intervals (P < 0.001). In multivariate analysis, microvascular invasion (mVI; P = 0.034) and ICGR15 ≥ 16% (P = 0.010) were independent risk factors for ERAH. In the ICGR1<16% subgroup, positive L3-AFP (P = 0.04), tumor size ≥5 cm (P = 0.011), surgical margin = 0 (P = 0.0103), mVI (P = 0.034), and extrahepatic recurrence were significant predictors of ERAH; in the ICGR15 ≥ 16%, subgroup, multiple tumors (P = 0.046) were identified as a risk factor for ERAH; however, this group did not experience much extrahepatic recurrence. ERAH was associated with mVI and ICGR15 ≥ 16%. Recurrence patterns and risk factors vary by liver function status, which should be considered in forming management strategies for early recurrence of HCC after curative hepatectomy.

In 320 living related liver transplantation performed between June 1990 and September 1997, there were 21 living related liver transplantation for patients with intrapulmonary shunting, manifested by digital clubbing, cyanosis, and dyspnea. We report the long-term outcome for more 6 months and our strategy to overcome complications in these recipients.A total of 21 patients (age range 2-33 years, 19 children and 2 adults, 6 males and 15 females) were classified into three grades according to shunt ratio calculated by TcMAA pulmonary scintigraphy; 5 in mild group (shunt ratio: less than 20%), 6 in moderated group (20%-40%), and 10 in severe group (more than 40%). The original underlying liver disease was biliary atresia in all patients.Spearmen's correlation coefficient rank test revealed that shunt ratio correlated significantly with PaO2 in room air (P=0.0001), PaO2 in 100% oxygen (P=0.0004), hematocrit (P=0.0276), and period of dyspnea before transplantation (P=0.023).Wound infection occurred in 80, 66, and 80%, and bile leakage in 20, 0, 40% in mild, moderate, and severe group, respectively. Patients who had portal vein thrombosis, and intracranial complication were classified as severe group and the incidence was 20 and 20%, respectively. The patient actuarial one year survival was 80, 66.7, and 48%, in mild, moderate, and severe group, respectively, although there was no significant difference. All patients who survived improved hepatopulmonary syndrome and the length of period required for the resolution was significantly correlated to the preoperative shunt ratio (P=0.023).Patients with severe shunting are susceptible to wound infection and bile leak. The trend of higher incidence of portal thrombosis and intracranial complications in the severe group was closely related high hematocrit. Secure surgical technique to reduce bile leak and delayed primary wound closure to reduce wound infection were found to be effective. Anticoagulant therapy by infusing heparin through the portal vein followed by coumadin could prevent fatal portal vein thrombosis without counter risk of fatal cerebral hemorrhage.

The expression of the genes for somatostatin (SRIF) and brain-derived neurotrophic factor (BDNF) was investigated in the central nervous system (CNS) of the macaque monkey (Macaca fuscata fuscata). Using Northern blot analysis, one SRIF mRNA transcript, 0.65 kb, and two BDNF mRNA transcripts, 1.6 and 4.0 kb in length, were detected in the monkey brain tissues. During the aging process (2 years, 10 years, and > 30 years), the ratio of SRIF mRNA/glyceraldehyde-3 phosphate dehydrogenase (G3PDH) mRNA significantly decreased (60-70%) in the hippocampus and in several cerebral subdivisions such as frontal cortex, temporal cortex, motor cortex, somatosensory cortex and visual cortex. BDNF mRNA was expressed in the various cerebral subdivisions and in the hippocampus. During the aging process, the gene expression of BDNF declined (20-50% for the 4.0 kb transcript, and 40-70% for the 1.6 kb transcript) in the various cerebral subdivisions. In the hippocampus, the level of the 1.6 kb mRNA at > 30 years old declined to 60% of the level at 2 years old, while the 4.0 kb mRNA did not change significantly during the aging process. Recent studies have shown that BDNF enhances the expression of SRIF mRNA in the rodent cerebral cortex (Nawa, H. et al., J. Neurochem., 60 (1993) 772-775; Nawa, H. et al., J. Neurosci., 14 (1994) 3751-3765). These studies and our present results suggest that the decrease in gene expression for a neurotrophic molecule, such as BDNF, might cause the levels of SRIF mRNA to decline in the primate brain during the aging process.

Through tropo-myosine-related kinase B (TrkB) receptors, brain-derived neurotrophic factor (BDNF) performs many biological functions such as neural survival, differentiation, and plasticity. T1, an isoform of TrkB receptors that lacks a tyrosine kinase, predominates in the adult mammalian CNS, yet its role remains controversial. In this study, to examine whether T1 transduces a signal and to determine its function, we first performed an affinity purification of T1-binding protein with the T1-specific C-terminal peptide and identified Rho GDP dissociation inhibitor 1 (GDI1), a GDP dissociation inhibitor of Rho small G-proteins, as a signaling protein directly associated with T1. The binding of BDNF to T1 caused Rho GDI1 to dissociate from the C-terminal tail of T1. Astrocytes cultured for 30 d expressed only endogenous T1 among the BDNF receptors. In 30 d cultured astrocytes, Rho GDI1, when dissociated in a BDNF-dependent manner, controlled the activities of the Rho GTPases, which resulted in rapid changes in astrocytic morphology. Furthermore, using 2 d cultured astrocytes that were transfected with T1, a T1 deletion mutant, or cyan fluorescent protein fusion protein of the T1-specific C-terminal sequence, we demonstrated that T1-Rho GDI1 signaling was indispensable for regulating the activities of Rho GTPases and for the subsequent morphological changes among astrocytes. Therefore, these findings indicate that the T1 signaling cascade can alter astrocytic morphology via regulation of Rho GTPase activity.

To investigate the effects of postlesion training on motor recovery, we compared the motor recovery of macaque monkeys that had received intensive motor training with those that received no training after a lesion of the primary motor cortex (M1). An ibotenic acid lesion in the M1 digit area resulted in impairment of hand function, with complete loss of digit movement. In the monkeys that had undergone intensive daily training (1 h/day, 5 days/wk) after the lesion, behavioral indexes used to evaluate manual dexterity recovered to the same level as in the prelesion period after 1 or 2 mo of postlesion training period. Relatively independent digit movements, including precision grip (prehension of a small object with finger-to-thumb opposition), were restored in the trained monkeys. Although the behavioral indexes of manual dexterity recovered to some extent in the monkeys without the postlesion training, they remained lower than those in the prelesion period until several months after M1 lesion. The untrained monkeys frequently used alternate grip strategies to grasp a small object with the affected hand, holding food pellets between the tip of the index finger and the dorsum of the thumb. These results suggest that the recovery after M1 lesion includes both use-dependent and use-independent processes and that the recovery of precision grip can be promoted by intensive use of the affected hand in postlesion training.

This study compared postoperative pain following four-port laparoscopic cholecystectomy (LC) and single-port cholecystectomy (SPC).This prospective, quasi-randomized, single-centre trial focusing on postoperative pain included 49 patients undergoing elective surgery with either a conventional LC, or SPC using a surgical glove port. Postoperative pain was evaluated using a visual analogue scale (VAS) and postoperative analgesic use as primary outcome measures. Total duration of operation, length of hospital stay, blood test results on the day after surgery and total port cost were secondary outcome measures.Twenty-five LCs and 24 SPCs were undertaken. The VAS score on day 1 after surgery was significantly less in the SPC group than in the LC group: median (range) 24 (12-38) versus 45 (33-57) mm (P = 0·002). Significantly fewer patients in the SPC group required analgesia (9 of 24 versus 19 of 25 in the LC group; P = 0·007). There were no significant differences in total duration of operation, length of hospital stay, and blood test results on the day after surgery.Single-port surgery using a surgical glove port reduces postoperative pain compared with conventional LC.

Hepatectomy is recommended as the most effective therapy for liver metastasis from colorectal cancer (CRCLM). It is crucial to elucidate the prognostic clinicopathological factors.Eighty-three patients undergoing initial hepatectomy for CRCLM were retrospectively analyzed with respect to characteristics of primary colorectal and metastatic hepatic tumors, operation details and prognosis.The overall 5-year survival rate after initial hepatectomy for CRCLM was 57.5%, and the median survival time was 25 months. Univariate analysis clarified that the significant prognostic factors for poor survival were depth of primary colorectal cancer (≥ serosal invasion), hepatic resection margin (< 5 mm), presence of portal vein invasion of CRCLM, and the presence of intra- and extrahepatic recurrence. Multivariate analysis indicated the presence of intra- and extrahepatic recurrence as independent predictive factors for poor prognosis. Risk factors for intrahepatic recurrence were resection margin (< 5 mm) of CRCLM, while no risk factors for extrahepatic recurrence were noted. In the subgroup with synchronous CRCLM, the combination of surgery and adjuvant chemotherapy controlled intrahepatic recurrence and improved the prognosis significantly.Optimal surgical strategies in conjunction with effective chemotherapeutic regimens need to be established in patients with risk factors for recurrence and poor outcomes as listed above.

Chronic stress is closely linked to the metabolic syndrome, diabetes, hyperuricemia and thromboembolism, but the mechanisms remain elusive. We reported recently that stress targets visceral adipose tissue (VAT), inducing lipolysis, low-grade inflammation with production of inflammatory adipokines, metabolic derangements such as insulin resistance, and prothrombotic state. In the present study, we hypothesized the involvement of VAT xanthine oxidoreductase (XOR), a source of reactive oxygen species (ROS) and uric acid (UA) in the above processes. Restraint stress in mice resulted in upregulation of XOR and xanthine oxidase activity, accumulation of ROS in VAT as well as liver and intestine, increase in serum UA levels, upregulation of NADPH oxidase subunits and downregulation of antioxidant enzymes. Immunohistochemistry and RT-PCR analysis also showed that restraint stress induced VAT monocyte accumulation and proinflammatory adipokine production, resulting in reduced insulin sensitivity and induction of plasminogen activator inhibitor-1 and tissue factor in VAT. Treatment with febuxostat, a potent XO inhibitor, suppressed stress-induced ROS production and VAT inflammation, resulting in improvement of serum UA levels, insulin sensitivity, and prothrombotic tendency. Our results suggest that stress perturbs glucose and UA metabolism, and promotes prothrombotic status, and that XO inhibition by febuxostat might be a potential therapy for stress-related disorders.

Abstract Living donor-morbidity was evaluated in 470 consecutive cases of living donor liver transplantation carried out from June 1990 to May 1999 at Kyoto University. Grafting was categorized into 4 groups according to the resection lines; left lateral segmentectomy (S2 + 3, n= 282, R1), extended left lateral segmentectomy without middle hepatic vein (MHV) (S2 + 3 + part4, n= 45, R2), left lobectomy with MHV (S2 + 3 + 4, n= 99, R3) and right lobectomy without MHV (S5 + 6 + 7 + 8, n= 43, R4). Intraoperative blood loss and operation duration were less for left lateral segmentectomy, but no significant difference was observed between left lobectomy and right lobectomy. The length of postoperative hospital stays was comparable among all groups except for the group with right lobe grafting. The AST values at the peak and at POD 7 were significantly elevated for right lobectomy, but the AST value normalized within one month in the majority of the cases. The close follow-up of donors with more than 1000 ml intraoperative bleeding, and of those donors who stayed in hospital for more than 30 days, the close follow-up, furthermore, of those donors with AST values higher than 100 IU/L AST after one month, revealed complete recovery. Biliary leakage was the most common and annoying complication after donor operations, especially in for right lobe grafting, but all donors recovered completely with conservative or minimal invasive therapy. The two cases of re-operation due to adhesive mechanical ileus we encountered were resolved completely. Finally, no donor-operation related death was noted. In conclusion, the morbidity of living donors is low or minimal even for right lobectomy, the most extended procedure, and complete recovery can be expected in all cases.

In countries where living donors are the only source of liver grafts, restrictions on graft size are a serious obstacle for the expansion of indications for adult recipients. To overcome this problem, auxiliary partial orthotopic liver transplants (APOLT*) was performed on the basis of the concept that the residual native liver would support the graft function until the graft had grown enough to function by itself.APOLT as an aid for small-for-size (SFS) grafts was reviewed retrospectively to evaluate its feasibility. Between April 1995 and March 1998, 20 recipients underwent APOLT, which was indicated because of a SFS graft in 15 of them. The indication was based on the estimated graft/recipient's body weight ratio (GRWR). If the ratio was <0.8%, APOLT was performed. The other 5 patients had a graft with a GRWR >0.8% and underwent APOLT on the basis of the residual native liver supporting the graft function temporarily, 4 for supplementation of the defective enzyme in metabolic liver diseases and one for leaving the potential of the regeneration of the native liver in fulminant hepatic failure. The recipients who underwent APOLT because of a SFS graft were categorized as the SFS group, and the others were the second group.In the SFS group, the age of the recipients ranged from 13 to 48 (median 23). The original indications of this group were fulminant hepatic failure in 2 recipients, acute deterioration of chronic liver diseases in 3, Wilson's disease in 2, biliary atresia in 4, primary biliary cirrhosis in 3, and primary sclerosing cholangitis (PSC) in one. The actual GRWR ranged from 0.45 to 0.72 (median 0.55). The graft was implanted after resection of the left lateral segment of the native liver. Except in the first two patients, the portal vein to the residual native liver was completely transected so that all of the portal blood drained into the graft liver. This procedure was successful in 9 patients. The cause of death in the other 6 was mainly infection. The mortality rate among the recipients with signs of advanced liver failure, such as massive ascites or hepatic coma, was higher, even though APOLT was used to support the SFS graft. In the second group, in the other five recipients who underwent APOLT for other indications, one recipient with fulminant hepatic failure died of sepsis caused by the dehiscence of bilio-enteric anastomosis.APOLT as an aid for a SFS graft is technically viable. This procedure can thus expand the indication of living donor liver transplants for adult recipients when the native liver retains some functional capability to support the grafted liver during the immediate postoperative period.

Autoimmune liver diseases (AILD) may progress to liver failure, requiring liver transplantation as definitive therapy, and these immune-mediated disorders may predispose the patient to more frequent graft rejection. The objective of this study was to determine the effect of preexisting AILD on the incidence of allograft rejection after liver transplantation. Sixty-three patients who underwent liver transplantation between March 1988 and December 1994 for AILDs that included autoimmune hepatitis (AIH; n = 33) and primary biliary cirrhosis (PBC; n = 30) were retrospectively compared with 47 patients who underwent liver transplantation for alcoholic cirrhosis during the same time period. There was a lower incidence of acute allograft rejection in patients with AILD who received tacrolimus-based compared with cyclosporine-based immunosuppression (50% v 85.5%; P = .02). However, patients with AILDs overall had a higher incidence of acute rejection than patients with alcoholic cirrhosis (81% v 46.8%; P < .001), regardless of the type of immunosuppression. In addition, steroid-resistant rejection occurred more frequently in patients with AILDs than in patients with alcoholic cirrhosis (38.1% v 12.8%; P = .003). There was also a trend toward a higher incidence of chronic rejection in patients with AILDs compared with patients with alcoholic cirrhosis (11.1% v 2.1%), but this difference did not reach statistical significance. Patient and graft survivals at 1 and 3 years were similar between patients with AILDs and alcoholic liver disease. Compared with alcoholic cirrhosis, preexisting AILDs are associated with a higher incidence of acute allograft rejection and a trend toward more frequent chronic rejection.

Abstract By using the developing monkey brain as a model for human development, we investigated the expression pattern of the FOXP2 gene, a member of the FOX family of transcription factors in the developing monkey brain, and compared its expression pattern with transcription factors PBX3, MEIS2 , and FOXP1 . We observed FOXP2 mRNA expression in several brain structures, including the striatum, the islands of Calleja and other basal forebrain regions, the cerebral cortex, and the thalamus. FOXP2 mRNA was preferentially expressed in striosomal compartments during striatal development. The striosomal expression was transient and developmentally down‐regulated in a topographical order. Specifically, during the perinatal state, striosomal FOXP2 expression was detected in both the caudate nucleus and the putamen, although expression was more prominent in the caudate nucleus than in the putamen. Striosomal FOXP2 expression declined during the postnatal period, first in the putamen and later in the caudate nucleus. During the same period, we also detected PBX3 mRNA in the striosomal compartment of the developing monkey striatum. FOXP2 , as well as PBX3 and MEIS2 , was expressed in the islands of Calleja and other cell clusters of the basal forebrain. FOXP2, in combination with PBX3 and MEIS2, may play a pivotal role in the development of striosomal neurons of the striatum and the islands of Calleja. J. Comp. Neurol. 509:180–189, 2008. © 2008 Wiley‐Liss, Inc.

Laparoscopic liver resection is currently performed in an increasing number of institutions as a minimally invasive treatment. However, no randomized controlled trials have compared laparoscopic and open liver resections. Twenty-three laparoscopic and 24 open liver resections for colorectal cancer liver metastases (CRCLM) were performed, and these data for both were retrospectively compared in the short-term results. The estimated blood loss was 99 ± 207 mL in the laparoscopic group and 397 ± 381 mL in the open group (P = 0.0018); blood loss was significantly higher in the open group. There were no differences in the surgical procedure, blood loss, transfusion rate, pathological margins, postoperative complications, 30-day mortality, duration of intravenous drip, or hospital stay. On postoperative courses, the values of total bilirubin, white blood cell count, and C-reactive protein were significantly lower in the laparoscopic group. The data of the present series suggest the lesser invasiveness and safety of laparoscopic liver resection even for patients with CRCLM, and they showed that postoperative laboratory tests were better after laparoscopy than after the traditional open approach with better short-term results. Tumor diameter less than 5 cm appears to be the appropriate indication for laparoscopic liver resection for CRCLM.

As the obesity epidemic worsens, the prevalence of fatty liver disease has increased. However, minimal data exist on the impact of combined fatty liver and metabolic syndrome on hepatectomy outcomes. Therefore, the aim of this analysis is to measure the outcomes of patients who do and do not have a fatty liver undergoing hepatectomy in the presence and absence of the metabolic syndrome.Patients with fatty and normal livers undergoing major hepatectomy (≥3 segments) were identified in the 2014 to 2018 American College of Surgeon National Surgical Quality Improvement Program database. Patients undergoing partial hepatectomy and those with missing liver texture data were excluded. Propensity matching was used and adjusted for multiple variables. A subgroup analysis stratified by the metabolic syndrome (body mass index ≥30 kg/m2, hypertension and diabetes) was performed. Demographics and outcomes were compared by χ2 and Mann-Whitney tests.Of 2,927 hepatectomies, 30% of patients (N = 863) had a fatty liver. The median body mass index was 28.6, and the metabolic syndrome was present in 6.3% of patients (N = 184). After propensity matching, 863 patients with fatty and 863 with normal livers were compared. Multiple outcomes were significantly worse in patients with fatty livers (P <.05), including serious morbidity (32% vs 24%), postoperative invasive biliary procedures (15% vs 10%), organ space infections (11% vs 7.8%), and pulmonary complications. Patients with fatty livers and the metabolic syndrome had significantly increased postoperative cardiac arrests, pulmonary embolisms, and mortality (P < .05).Fatty liver disease is associated with significantly worse outcomes after major hepatectomy. The metabolic syndrome confers an increased risk of postoperative mortality.

Living donor liver transplantation (LDLT) was originally indicated only for elective cases of pediatric patients with end-stage liver disease. In Japan, however, where liver transplantation from brain-dead donor is performed very rarely, this indication has been expanded to emergency cases such as fulminant hepatic failure (FHF).Thirty-eight patients with FHF were treated between May 1992 and April 1999. Causes of acute liver failure were non-A, non-B hepatitis in 27 patients, hepatitis B virus in seven, and hepatitis A virus, Epstein-Barr virus, herpes simplex virus, and chrome poisoning in one each.Four patients did not undergo LDLT because of severe brain damage or combined multiple organ failure. The remaining 34 patients underwent a total of 36 LDLTs, including two retransplantations; 16 children received transplants of 17 lateral segments, three children and eight adults transplants of 11 left lobes, and seven adults transplants of eight right lobes. A total of 15 recipients died, four of primary graft dysfunction, three of refractory acute rejection, two of pneumonia, and one each of ductopenic rejection, sepsis, aplastic anemis, recurrence of Epstein-Barr virus hepatitis, multiple organ failure by chrome poisoning, and unknown hepatic failure. Primary graft dysfunction developed in adult recipients with small-for-size graft transplants, whereas refractory acute rejection and ductopenic rejection occurred in six grafts each of children with non-A, non-B FHF.LDLT can be safely expanded to cases of FHF in adult patients. Primary graft dysfunction in adult recipients with small-for-size left lobe grafts can be overcome by using right lobes. However, refractory acute rejection and ductopenic rejection in children remain a major problem.

We investigated the changes of brain-derived neurotrophic factor (BDNF)-immunoreactive structures in the hippocampal formation of aged macaques (Macaca fuscata fuscata). At adult stages (10 and 12 years), BDNF immunoreactivity occurred in the neurons of the dentate gyrus, the pyramidal neurons in the CA1, CA2, CA3 subfields and the subiculum, and the neurons in the CA4 subfield and the entorhinal cortex. The apical dendrites were also BDNF immunopositive. In aged monkeys (26, 30 and 32 years), the intensity of the BDNF-immunoreactivity declined significantly in cell bodies and dendrites of the neurons in the hippocampal formation except the CA2 pyramidal neurons. These findings indicate that BDNF is one of the vulnerable signal molecules during the aging process of the primate hippocampal formation.

Clinical experience suggests that grafts obtained from steatotic livers result in primary nonfunction more frequently than those from nonsteatotic livers. To date, however, only a few studies have been done to verify the accuracy of this observation. To investigate the effects of cold preservation on steatotic liver viability, liver grafts obtained from rats fed with a choline-deficient diet were transplanted after periods of cold preservation. Recipient survival rates with normal liver grafts were 8/8 (100%) and those with steatotic liver grafts were 7/8 (88%) (P > 0.05) after 1-hr preservation with UW solution. After 9-hr preservation, however, these rates decreased significantly to 0/8 (0%) with steatotic grafts (P < 0.01), but were not significantly decreased with normal grafts. LDH levels in the effluent at the time of transplantation were 133 IU/L (1-hr) and 512 IU/L (9-hr) in normal livers, but in steatotic livers these were elevated to 598 and 3141 IU/L, respectively (P < 0.01). Recovery rates of hepatic blood flow measured by laser Doppler flowmeter after revascularization were 99% (1-hr) and 96% (9-hr) in normal grafts, but in steatotic grafts they were 98% (1-hr) and 63% (9-hr, P < 0.01). In addition, the oxidative phosphorylation ability of liver mitochondria obtained from steatotic grafts was decreased significantly after cold preservation. The present results suggest that steatotic liver grafts are prone to lose their viability more easily than normal liver grafts after prolonged periods of cold preservation due to a combination of causes.

By altering their morphology, astrocytes, including those involved in the maintenance and plasticity of neurons and in clearance of transmitter, play important roles in synaptic transmission; however, the mechanism that regulates the morphological plasticity of astrocytes remains unclear. Recently, we reported that T1, a subtype of TrkB (a family of BDNF-specific receptors), altered astrocytic morphology through the control of Rho GTPases in primary astrocyte cultures. In this study, we extended this observation to investigate acute neocortical slices from adult rats. T1 siRNA-expression vectors were electroporated into astrocytes in neocortical layer I of living rats. In both normal slices and control vector-electroporated slices, BDNF induced the elongation of the astrocytic processes and increased the branching of processes in slices after 1 h incubation. In contrast, in T1 siRNA-electroporated slices, no such significant morphological changes were observed in the astrocytes. In addition, the number of synaptophysin+ sites in contact with GFAP+ processes increased in a BDNF-T1-dependent manner without the increase in the total synaptophysin+ sites. Therefore, the present study provides evidence of the regulation of layer I astrocytic morphology by the BDNF-T1 signal in adult rat neocortical slices.

A large-scale purification of monkey brain arylamidase was carried out. Amino acid analyses indicate that the enzyme is rich in acidic amino acids and is poor in cystine. The amino terminal residue was determined to be alanine by dansylation. The enzyme was activated by sulfhydryl compounds. Dithiothreitol was more effective than β-mercaptoethanol. Bestatin competitively inhibited the enzyme activity and the Ki value was calculated to be 2.5×10−7 M, which was of the same order as that of puromycin. The inhibitions by puro-mycin and bestatin were reversible. The enzyme hydrolyzed di-, tri-, and ohgopeptides including physiologically active peptides. Of physiologically active peptides, enkephalins and Met-Lys-bradykinin, which possess a neutral amino acid at the N-terminal position, were more rapidly hydrolyzed by the enzyme. Peptides such as LH-RH and TRH, which possess a pyrrolidonecarboxylyl group at the N-terminal position, and substance P and bradykinin, which possess a proline residue adjacent to the N-terminal residue, were not hydrolyzed by the enzyme. The Km values for various peptides indicate that the enzyme has higher affinity for oligopeptides than di- and tripeptides. The aminopeptidase activity of the enzyme was also competitively inhibited by puromycin and bestatin. Analyses of the hydrolysis products of various peptides by the dansylation method indicate that the enzyme has both kinin-converting activity and angiotensinase activity.

Recently, the truncated TrkB receptor, T1, has been reported to be involved in the control of cell morphology via the regulation of Rho proteins, through which T1 binds Rho guanine nucleotide dissociation inhibitor (Rho GDI) 1 and dissociates it in a brain-derived neurotrophic factor (BDNF)-dependent manner. However, it is unclear whether T1 signaling regulates the downstream of Rho signaling and the actin cytoskeleton. In this study, we investigated this question using C6 rat glioma cells, which express T1 endogenously. Rho GDI1 was dissociated from T1 in a BDNF-dependent manner, which also causes decreases in the activities of Rho-signaling molecules such as RhoA, Rho-associated kinase, p21-activated kinase, and extracellular-signal regulated kinase1/2. Moreover, BDNF treatment resulted in the disappearance of stress fibers in the cells treated with lysophosphatidic acid, an activator of RhoA, and in morphological changes in cells. Furthermore, a competitive assay with cyan fluorescent protein fusion proteins of T1-specific sequences reduced the effects of BDNF. These results suggest that T1 regulates the Rho-signaling pathways and the actin cytoskeleton.

Among a panel of monoclonal antibodies generated against monkey brain tissue, a class of antibodies was found to produce perineuronal staining of small subsets of mammalian central neurons. Three antibodies (MAbs 473, 376, 528) we report here define two different, though partially overlapping, neuronal subsets in the monkey neocortex. All 3 antibodies stain in addition certain chondrocytes. The neural immunoreactivities were lost, and the chondral immunoreactivities either lost or enhanced, after treatment of the sections with chondroitinase ABC. Independently, 3 other antibodies (MAbs 1B5, 9A2, 3B3) with established specificity to glycosaminoglycan epitopes also produced perineuronal staining of a related subset of central neurons. Immuoblot experiments with two of the antibodies revealed bands of high molecular weight. These findings indicate that certain glycosaminoglycans occur surrounding mammalian central neurons, and suggest that different neuronal subsets are associated with different combinations of proteoglycan epitopes.

Antisera to substance P, somatostatin and vasoactive intestinal polypeptide were produced in rabbits and used to establish radioimmunoassays. The specificities and sensitivities of the assays were characterised before using them to measure the levels of these peptides in the paravertebral sympathetic ganglia and spinal sensory ganglia of embryonic chicks. The immunoreactive materials in the developing ganglia were shown to be identical with the peptides by the criterion of co-elution on reverse-phase high performance liquid chromatography. The levels of the peptides varied independently during the developmental period studied between day 8 and hatching at day 20 of incubation. Vasoactive intestinal polypeptide was first detectable on day 12 in the sympathetic ganglia and the levels increased through to hatching, at which time it first became detectable in the sensory ganglia. In contrast, substance P and somatostatin were detectable in both sensory and sympathetic ganglia throughout the period studied: the content of substance P increased up to day 18 after which time it declined, and somatostatin was present initially (day 8) at high levels before rapidly declining to a minimum at day 10 and then slowly increasing again, in parallel with the general growth of the ganglia, reflected by their protein contents. The differential changes in levels of substance P, somatostatin and vasoactive intestinal polypeptide during development indicates that different regulatory mechanisms are responsible for the ontogenesis of individual peptide phenotypes in the peripheral nervous system.

The intercalated cell masses of the amygdala consist of cell clusters located between the basolateral complex of the amygdala and its surrounding structures including the central nucleus of the amygdala and the external capsule. Although recent studies have revealed that the intercalated cell masses may play an important role in emotional learning and memory, there are only a few reports on its molecular characterization. We examined the expression patterns of transcription factors in the intercalated cell masses in late embryonic stage and postnatal rats, and non-human primates. Dlx5, Foxp2, Pbx3 and Meis2 were expressed in all subdivisions of the intercalated cell masses, while Ebf1, Nkx2.1 and Foxp1 were not. In contrast, Pax6 was only expressed in a small population of the main intercalated islands, but not in the medial or lateral cell clusters. In addition, few Pax6-positive neurons co-expressed Foxp2. Thus the intercalated cell masses do not contain a homogeneous population of neurons, in terms of their molecular constituents. Given that Foxp2, Pbx3 and Meis2 are preferentially expressed in distinct cell populations in the developing striatum, and that the intercalated cell masses of the amygdala appear to be a ventrocaudal expansion of the striatum, the intercalated neurons may share a common origin with some types of neurons located in the dorsal striatum.

Background The VIO soft-coagulation system (SCS) is a new device for tissue coagulation. The current study evaluated the efficacy of the SCS when used for liver resection. Methods The 252 patients were divided into 2 groups; in 155 patients (conventional group), liver transection was performed using an ultrasonic dissector and saline-coupled bipolar electrocautery for hemostasis. In 97 patients (SCS group), the SCS was used instead of bipolar electrocautery. Results The median blood loss and surgical time were less in the SCS group than in the conventional group (350 vs 640 mL, P = .0028; 280 vs 398 min, P < .0001). No significant differences were found in postoperative complications between the SCS group (32.0%) and the conventional group (40.6%). The risk factors for bleeding were nonuse of the SCS (P = .0039), macroscopic vascular invasion of the hepatic tumors (P = .0088), and collagen type IV value in the sera >200 (P = .0250) on multivariate analysis. In a subgroup analysis, in the collagen type IV value >200 subgroup, the tumor diameter >5 cm subgroup, and the inflow nonocclusion subgroup, use of the SCS decreased surgical bleeding (P = .0120, P = .0126, and P = .0032, respectively) and surgical time (P = .0001, P < .0001, and P = .0036, respectively) compared with the conventional group. Furthermore, even in the major hepatectomy group, the SCS use decreased surgical time (P < .0001). Conclusion The SCS is an effective and safe device for decreasing surgical time and surgical bleeding without increasing the rate of bile leakage and causing other complications. The VIO soft-coagulation system (SCS) is a new device for tissue coagulation. The current study evaluated the efficacy of the SCS when used for liver resection. The 252 patients were divided into 2 groups; in 155 patients (conventional group), liver transection was performed using an ultrasonic dissector and saline-coupled bipolar electrocautery for hemostasis. In 97 patients (SCS group), the SCS was used instead of bipolar electrocautery. The median blood loss and surgical time were less in the SCS group than in the conventional group (350 vs 640 mL, P = .0028; 280 vs 398 min, P < .0001). No significant differences were found in postoperative complications between the SCS group (32.0%) and the conventional group (40.6%). The risk factors for bleeding were nonuse of the SCS (P = .0039), macroscopic vascular invasion of the hepatic tumors (P = .0088), and collagen type IV value in the sera >200 (P = .0250) on multivariate analysis. In a subgroup analysis, in the collagen type IV value >200 subgroup, the tumor diameter >5 cm subgroup, and the inflow nonocclusion subgroup, use of the SCS decreased surgical bleeding (P = .0120, P = .0126, and P = .0032, respectively) and surgical time (P = .0001, P < .0001, and P = .0036, respectively) compared with the conventional group. Furthermore, even in the major hepatectomy group, the SCS use decreased surgical time (P < .0001). The SCS is an effective and safe device for decreasing surgical time and surgical bleeding without increasing the rate of bile leakage and causing other complications.

Antibiotic prophylaxis is frequently administered after liver resection to prevent postoperative infections. However, very few studies have examined the usefulness of antibiotic prophylaxis after liver resection. A randomized controlled trial was conducted to evaluate the postoperative antibiotic prophylaxis in patients after liver resection.A total of 241 patients scheduled to undergo liver resection were randomly assigned to the non-postoperative antibiotic group (n = 95) or the antibiotic group (n = 95). The antibiotic group was given flomoxef sodium every 12 hours for 3 days after the operation. The end point was signs of infection, surgical site infection, or infectious complications.There were no significant differences between the 2 groups in signs of infection (21.3% vs 25.5%, P = .606), the incidence of systemic inflammatory response syndrome (11.7% vs 17.0%, P = .406), infectious complications (7.5% vs 17.0%, P = .073), surgical site infection (10.6% vs 13.8%, P = .657), and remote site infection (2.1% vs 8.5%, P = .100).Postoperative antibiotic prophylaxis cannot prevent postoperative infections after liver resection, and it is thought that antibiotic prophylaxis is unnecessary and costly.

Using receptor subunit-specific antibodies, the cellular localization of NMDA and AMPA type glutamate receptor subunits was studied within the rodent (rat, guinea pig) and non-human primate (monkey) inner ear. In the spiral and vestibular ganglion, almost all cells were immunoreactive for the NMDAR1 subunit and the AMPA type receptor subunit GluR2/3. This indicates that both NMDA and non-NMDA type glutamate receptors may be co-distributed in the primary afferent neuronal components, and are possibly involved in neurotransmission in the primary auditory and vestibular systems. This study also indicated the possible localizations of glutamate receptors in the nonneuronal cells in the inner ear, suggesting that some nonneuronal cells may also have the ability to mediate glutamate signalling.

Cao S, Cox KL, Berquist W, Hayashi M, Concepcion W, Hammes GB, Ojogho OK, So SKS, Frerker M, Castillo RO, Monge H, Esquivel CO. Long‐term outcomes in pediatric liver recipients: Comparison between cyclosporin A and tacrolimus. Pediatr Transplantation 1999: 3: 22–26. © Munksgaard, 1999 In recent years, Tacrolimus ® (FK506, TAC) has been increasingly utilized in liver transplantation. However, long‐term risks and benefits as compared with conventional cyclosporin A (CsA) have not been fully elucidated. This retrospective study examined the potential outcome differences between TAC‐ and CsA‐based immunosuppressive therapy in pediatric liver transplant recipients. From March 1988 to December 1996, 218 children (aged 0.1 – 17 yr) underwent 238 orthotopic liver transplantations; 58.7% (128/218) were under 2 yr old at time of transplant. Initially, the maintenance immunosuppressive regimen consisted of CsA and prednisone, with antilymphocytic preparations (MALG, ATGAM, and OKT3) as induction therapy. Subsequently, TAC was used first as rescue therapy for steroid refractory rejection in CsA patients and then as maintenance immunosuppression. Fifty‐seven out of the 147 CsA patients were converted to TAC for various reasons while 71 patients were placed on TAC as primary maintenance immunosuppression. 62.6 per cent (92/147) of liver recipients on CsA experienced at least one biopsy‐proven acute rejection episode as compared to 50.7% (36/71) for TAC patients (p = 0.09); likewise, 34% (50/147) of CsA patients had more than one episode of rejection vs. 18.3% (13/71) for patients on TAC (p &lt; 0.02). Rejection was the reason for conversion from CsA to TAC in 29 of 57 patients. Conversely, 19.0% (28/147) of CsA patients had to be switched to TAC for reasons not related to rejection (i.e. side‐effects). The overall incidence of histologically proven chronic rejection was 7.8% (17/218). 10.9 per cent (16/147) of the children who were on CsA initially developed chronic rejection, which was significantly higher compared with one of 71 TAC recipients (p &lt; 0.02). Of these 16 CsA patients with chronic rejection, 50.0% (8/16) underwent retransplantation for graft failure (mean interval from time of diagnosis of chronic rejection to re‐transplant, 4.0 months; range 1–8 months), whereas the TAC patient has remained clinically stable with normal LFTs after 23 months of follow‐up. One year after liver transplantation, 72.8% (107/147) of CsA patients were still on steroids (mean dosage 0.20 mg/kg/d), as compared to 42.3% (30/71) of the TAC patients (mean dosage 0.14 mg/kg/d). The incidence of post‐transplant lymphoproliferative disorder (PTLD) in Epstein–Barr virus (EBV)‐infected patients was 2.2% (2/90), 7.0% (5/71) and 12.3% (7/57) for CsA, primary and TAC‐converted groups, respectively. The overall incidence of PTLD was 6.9% (15/218). In summary, pediatric liver transplant recipients treated with TAC as primary maintenance immunosuppressive medication experienced significantly fewer episodes of rejection; especially chronic rejection, which lead to graft loss. However, the trade‐off is a potential increased incidence of EBV‐related PTLD in these patients.

The concentrations of vasoactive intestinal polypeptide, somatostatin and substance P were determined in various cerebral subdivisions of adult and foetal Japanese monkeys (Macaca fuscata fuscata) by specific radioimmunoassays. In adult tissues, the highest level of vasoactive intestinal polypeptide was found in the somatosensory cortex and the lowest level in the occipital cortex. A high level of somatostatin was found in the association cortex (prefrontal, parietal and temporal cortex); the lowest level was noted in the occipital cortex. Substance P was found to be high in prefrontal and temporal cortex. The highest levels of substance P and somatostatin were obtained in the amygdala. Between embryonic 4 and 5.5 months, concentrations of peptides increased dramatically, and in the adult, all neuropeptides in cortical subdivisions significantly decreased. By the gel filtration method, only one immunoreactivity which coeluted with substance P and vasoactive intestinal polypeptide was demonstrated in extracts of 4-, 5.5-month-old and adult monkey cerebral cortex. In contrast, somatostatin immunoreactivity eluted as 3 peaks. Almost 80% of the immunoreactivity co-eluted with synthetic somatostatin, regardless of the age of the tissue. The molecular weights of two larger molecules were determined to be 13 and 3 kdaltons.

Abstract Background and Study Aim The present study assesses the feasibility as well as the technical and functional success rates of a novel endoscopic ultrasound‐guided hepaticogastrostomy ( EUS‐HGS ) technique called the locking stent method that uses end‐bare covered metallic stents ( EBCMS ). Methods Twenty consecutive patients who were histologically diagnosed with unresectable cancer complicated with obstructive jaundice underwent EUS‐HGS due to failed endoscopic biliary drainage or inaccessible papilla. We retrospectively collected clinical data for these patients including technical and functional success rates and complications. Results Seven were treated by EUS‐HGS ( EUS‐HGS group), and 13 were treated using the locking stent EUS‐HGS method ( LS group). Technical and functional success rates were 100% in both groups. Procedural duration did not significantly differ between the EUS‐HGS and LS groups (26.9 ± 9.0 versus 32.3 ± 11.1 min, P = 0.30). Two patients developed complications related to stent migration in the EUS‐HGS group. In contrast, although mild post‐procedural bile peritonitis required conservative treatment for a few days, none of the stents malfunctioned in the LS group. Conclusion Our method can safely and effectively prevent stent dysfunction, but validation in a prospective clinical trial is required.

Background The benefits of anatomic resection in patients with small (<5 cm), solitary hepatocellular carcinomas remain unclear. Outcomes were therefore evaluated in patients who underwent anatomic resection or nonanatomic resection of small solitary hepatocellular carcinomas. Methods Factors affecting overall survival and disease-free survival were investigated in 330 patients who underwent curative hepatectomy for solitary (≤5 cm) hepatocellular carcinomas without macroscopic vascular invasion. In addition, a propensity score matching model with 330 patients was constructed to overcome bias, with subgroups analyzed by tumor diameter (<3 cm and 3–5 cm). Results ICG-R15 ≥25% was confirmed as being independently associated with poorer overall survival and disease-free survival. One-to-one matching of preoperative characteristics yielded 72 pairs of patients receiving anatomic resection and nonanatomic resection, with long-term outcomes, including overall survival and disease-free survival, being similar in these 2 groups. Subgroup analysis showed that, in patients with tumors <3 cm in diameter, short-term outcomes were better in the nonanatomic resection group than in the anatomic resection group, including significantly reduced operation time (P = .02), blood loss (P = .01), blood transfusion (P < .01), complications (particularly bile leakage and abdominal abscess) (P = .04), and postoperative hospital stay (P < .01). Conclusion Anatomic resection was not superior to nonanatomic resection in survival outcomes in patients with solitary small hepatocellular carcinomas without macroscopic vascular invasion. Rather, postoperative short-term outcomes were more favorable with nonanatomic resection. The benefits of anatomic resection in patients with small (<5 cm), solitary hepatocellular carcinomas remain unclear. Outcomes were therefore evaluated in patients who underwent anatomic resection or nonanatomic resection of small solitary hepatocellular carcinomas. Factors affecting overall survival and disease-free survival were investigated in 330 patients who underwent curative hepatectomy for solitary (≤5 cm) hepatocellular carcinomas without macroscopic vascular invasion. In addition, a propensity score matching model with 330 patients was constructed to overcome bias, with subgroups analyzed by tumor diameter (<3 cm and 3–5 cm). ICG-R15 ≥25% was confirmed as being independently associated with poorer overall survival and disease-free survival. One-to-one matching of preoperative characteristics yielded 72 pairs of patients receiving anatomic resection and nonanatomic resection, with long-term outcomes, including overall survival and disease-free survival, being similar in these 2 groups. Subgroup analysis showed that, in patients with tumors <3 cm in diameter, short-term outcomes were better in the nonanatomic resection group than in the anatomic resection group, including significantly reduced operation time (P = .02), blood loss (P = .01), blood transfusion (P < .01), complications (particularly bile leakage and abdominal abscess) (P = .04), and postoperative hospital stay (P < .01). Anatomic resection was not superior to nonanatomic resection in survival outcomes in patients with solitary small hepatocellular carcinomas without macroscopic vascular invasion. Rather, postoperative short-term outcomes were more favorable with nonanatomic resection.

Portal vein reconstruction is a crucial factor affecting the outcome of a successful living-related liver transplantation. We describe here our experience with portal vein reconstruction in 314 cases of living-related liver transplantation with use of novel surgical modalities to enable the transplant surgeons to deal with any size mismatch between the donor's and recipient's portal veins.Portal vein reconstruction was classified into 2 major groups, anastomosis without and with a vein graft. When there was no stenosis of the recipient portal vein and the diameter was the same, the portal trunk was used for anastomosis. When the diameter mismatch was minimal, branch patch anastomosis was feasible. When the recipient portal vein was significantly stenotic and the portal vein of the graft was long enough, we removed the stenotic trunk and constructed an anastomosis between the graft portal vein and the confluence of the recipient portal vein. When the graft portal vein was short, a vein graft was interposed. The vein patch technique was preferable when the diameter of the graft vein was not large enough for the interposition technique.Anastomosis without vein graft included trunk anastomosis (n = 156), branch patch anastomosis (n = 39), and confluence anastomosis (n = 22). Anastomosis with vein graft used the interposition technique (n = 77) and vein patch technique (n = 27). The origin of the grafts was mostly from the maternal left ovarian vein (70%) or the paternal inferior mesenteric vein (27%). Complications related to portal vein reconstruction occurred in 16 (5%) patients: portal vein thrombosis in 8, stenosis in 7, and fatal rupture in 1 patient. The incidence of complications was similar for all techniques except for confluence anastomosis.Our innovative techniques should be helpful for overcoming diameter or length mismatches in portal vein reconstruction in pediatric liver transplantation.

Antimitochondrial antibodies (AMA) which recognize the E2 component of the pyruvate dehydrogenase complex are found in virtually all patients with the autoimmune liver disease primary biliary cirrhosis (PBC). The factors that contribute to elevated AMA and the relationship of the autoantibodies to disease pathogenesis have not been elucidated. Since cytokines are important regulators of antibody production and isotype switching, the association of specific cytokines to antibody production was examined in patients with PBC. Elevations in IL-2, IFN-γ, IL-4, IL-5, and IL-10 were detected in serum from patients with PBC. However, only IFN-γ (6012 ± 1128 pg/ml vs 147 ± 89 pg/ml,P< 0.0001) and IL-5 (382 ± 103 pg/ml vs 29 ± 12 pg/ml,P< 0.001) were significantly elevated compared to normal controls. Moreover, there was a positive correlation in the levels of IFN-γ, and to a lesser extent IL-5, with the levels of soluble CD30 (sCD30) in the circulation. The elevated levels of sCD30 detected in patients with PBC (194 ± 29 U/ml vs 39 ± 9 U/ml in normal controls) suggest that CD30+cells may produce cytokines, which contribute to the immune abnormalities in patients with PBC.

Distribution of somatostatin-immunoreactive cells in the cerebral cortex of macaque monkeys at embryonic day 120 (E120), E140, newborn, postnatal day 60 (P60) and adult stages were studied by the avidin-biotin-peroxidase immunohistochemical method. At all stages, there existed 3 types of cells in the gray matter: bipolar, multipolar and small-sized cells which stained only in perikaryon. Somatostatin-immunoreactive cells were observed from E120. The cell number increased between E120 and E140 and decreased until P60. At the newborn stage, a high density of cells was distributed in layer II of the prefrontal and parietal cortices (areas FD and PE). In layer I of the postcentral, parietal, temporal and preoccipital cortices (areas FA, PC, PE, TA, TE and OA), small numbers of horizontal cells were detected only at the embryonic and newborn stages. In adulthood, the number of somatostatin cells was much smaller than at the early stages (E140 and newborn). Compared to other cortical areas, in occipital cortex (area OC), there was little change in cell number during development. In occipito-temporal cortices, there were increases in cell number from posterior to anterior portion at all the stages. The large number of somatostatin cells in all layers of the cerebral cortex during the early stages indicates that somatostatin plays a role in the development of the monkey cerebral cortex.

A major concern in adult-to-adult living donor liver transplantation is the selection of graft type; that is, is it is better to use the right lobe with or without the middle hepatic vein (MHV)? This choice has a considerable impact on donor safety, vascular reconstruction and graft function in the recipient. To facilitate making an appropriate choice, on the basis of a preliminary study (n = 17), we herein propose a graft selection algorithm using three parameters: graft-to-recipient body weight ratio (GRWR), percentage remnant liver volume (%RLV) and estimated congestion ratio (ECR). The algorithm was evaluated with 50 consecutive cases with respect to postoperative liver function of donors and recipients and survival of recipients. Postoperative recovery was comparable between the two groups (p = NS). The overall cumulative 18-month survival rate was 86.7% for the 'with MHV graft group', and 76.1% for the gwithout MHV graft grouph (p = NS). For 41 cases (82%), graft types were chosen according to the algorithm, whereas the remaining 9 cases (18%) needed detailed discussion of donor, recipient and operative factors. In conclusion, we constructed a graft selection algorithm based on congestion volume, which will contribute to objective graft-type selection in adult-to-adult LDLT.

The enzyme activities of glutamic acid decarboxylase (GAD), tyrosine hydroxylase (TH) and choline acetyltransferase (ChAT) and concentrations of substance P (SP) and somatostatin were determined in the cerebellum of macaque monkey (Macaca fuscata fuscata) at 3 different ages, embryonic 4 months, embryonic 5.5 months (full-term) and adult. Similar graded increases in the activities of GAD and TH were observed during development. In contrast, ChAT activity was relatively high at embryonic 4 months, increased about twofold between embryonic 4 months and 5.5 months, but did not change between embryonic 5.5 months and adult. These findings suggest that noradrenergic terminals develop synchronously with GABAergic interneurons. On the other hand, the innervation by ChAT-containing fibers is completed during the prenatal period. The concentrations of somatostatin and SP were high at embryonic 4 months, and decreased to, respectively, about 1/18 and 1/4 (expressed per g weight) in adult animals. Several interpretations of the decrease of the two neuropeptides in cerebellar tissue during ontogeny are discussed.

Hepatocellular carcinoma (HCC) shows a high rate of recurrence after hepatectomy; predictive factors for early recurrence would help determine optimal therapeutic and management strategies. Among 163 patients with HCC undergoing hepatectomy with curative intent, 46 patients developed recurrence within 1 year. Clinicopathological data were retrospectively analyzed to identify predictive parameters for early recurrence. Survival rates in cases of recurrence within 1 year were worse than those of no recurrence within 1 year or recurrence after 1 year. Protein induced by vitamin K absence/antagonist II (PIVKA-II) greater than 150, positive fucosylated alpha-fetoprotein (L3-AFP), and deviancy from Milan criteria (MC) on preoperative imaging were associated with high risk of early recurrence and total number of these three risk factors predicted the survival. With multivariate analysis, (1) preoperatively, positive factors of two or more among three items of PIVKA-II, L3-AFP, and deviancy from MC; (2) and postoperatively, pathological cancer spread (microscopic vascular invasion and/or intrahepatic metastasis) both represented risks for early recurrence. A combination of three preoperative factors, PIVKA-II, L3-AFP, and MC status, in conjunction with the postoperative factor of cancer spread status represents a significant indicator for recurrence within 1 year. Improving the prognosis of patients with HCC would depend on how to adequately treat those at high risk of early recurrence.

Hepatic pseudolymphoma (HPL) and primary hepatic marginal zone B cell lymphoma of mucosa-associated lymphoid tissue (MALT lymphoma) are rare diseases and the differential diagnosis between these two entities is sometimes difficult. We herein report a 56-year-old Japanese woman who was pointed out to have a space occupying lesion in the left lateral segment of the liver. Hepatitis viral-associated antigen/antibody was negative and liver function tests including lactic dehydrogenase, peripheral blood count, tumor markers and soluble interleukin-2 receptor were all within normal limit. Imaging study using computed tomography and magnetic resonance imaging were not typical for hepatocellular carcinoma, cholangiocarcinoma, or other metastatic cancer. Fluorodeoxyglucose-positron emission tomography examination integrated with computed tomography scanning showed high standardized uptake value in the solitary lesion in the liver. Under a diagnosis of primary liver neoplasm, laparoscopic-assisted lateral segmentectomy was performed. Liver tumor of maximal 1.0 cm in diameter was consisted of aggregation of lymphocytes of predominantly B-cell, containing multiple lymphocyte follicles positive for CD10 and bcl-2, consistent with a diagnosis of HPL rather than MALT lymphoma, although a definitive differentiation was pending. The background liver showed non-alcoholic fatty liver disease/early non-alcoholic steatohepatitis. The patient is currently doing well with no sign of relapse 13 months after the surgery. Since the accurate diagnosis is difficult, laparoscopic approach would provide a reasonable procedure of diagnostic and therapeutic advantage with minimal invasiveness for patients. Considering that the real nature of this entity remains unclear, vigilant follow-up of patient is essential.

A strong causal link exists between psychological stress and insulin resistance as well with hypertension. Meanwhile, stress-related responses play critical roles in glucose metabolism in hypertensive patients. As clinical trials suggest that angiotensin-receptor blocker delays the onset of diabetes in hypertensive patients, we investigated the effects of irbesartan on stress-induced adipose tissue inflammation and insulin resistance. C57BL/6J mice were subjected to 2-week intermittent restraint stress and orally treated with vehicle, 3 and 10 mg/kg/day irbesartan. The plasma concentrations of lipid and proinflammatory cytokines [Monocyte Chemoattractant Protein-1 (MCP-1), tumor necrosis factor-α, and interleukin-6] were assessed with enzyme-linked immunosorbent assay. Monocyte/macrophage accumulation in inguinal white adipose tissue (WAT) was observed with CD11b-positive cell counts and mRNA expressions of CD68 and F4/80 using immunohistochemistry and RT-PCR methods respectively. The mRNA levels of angiotensinogen, proinflammatory cytokines shown above, and adiponectin in WAT were also assessed with RT-PCR method. Glucose metabolism was assessed by glucose tolerance tests (GTTs) and insulin tolerance tests, and mRNA expression of insulin receptor substrate-1 (IRS-1) and glucose transporter 4 (GLUT4) in WAT. Restraint stress increased monocyte accumulation, plasma free fatty acids, expression of angiotensinogen and proinflammatory cytokines including MCP-1, and reduced adiponectin. Irbesartan reduced stress-induced monocyte accumulation in WAT in a dose dependent manner. Irbesartan treatment also suppressed induction of adipose angiotensinogen and proinflammatory cytokines in WAT and blood, and reversed changes in adiponectin expression. Notably, irbesartan suppressed stress-induced reduction in adipose tissue weight and free fatty acid release, and improved insulin tolerance with restoration of IRS-1 and GLUT4 mRNA expressions in WAT. The results indicate that irbesartan improves stress-induced adipose tissue inflammation and insulin resistance. Our results suggests that irbesartan treatment exerts additive benefits for glucose metabolism in hypertensive patients with mental stress.

Background Postoperative bile leakage can be a serious complication after hepatic resection. Few studies have analyzed patients according to the time of onset of bile leakage. We analyzed differences between patients with early- and late-onset bile leakage after hepatic resection and assessed clinical characteristics and outcomes in patients with late-onset leakage. Methods Between 2008 and 2010, 1,009 patients underwent hepatic resection at 4 participating university hospitals and 2 community hospitals. Fifty-two patients (5.1%) with postoperative bile leakage were divided into an early-onset group (<2 weeks after surgery, n = 34) and a late-onset group (≥2 weeks after surgery, n = 18). Patient characteristics and outcomes were collected prospectively and analyzed retrospectively. Results The proportion of patients who underwent intra-abdominal placement of a drainage catheter was significantly less in the late-onset group than the early-onset group. All 18 patients in the late-onset group developed intra-abdominal infection, and 2 died of sepsis. The proportion of patients who underwent invasive treatment (abdominal paracentesis, endoscopic biliary drainage, or second hepatic resection) was significantly greater in the late-onset group than in the early-onset group. The time to resolution of bile leakage was significantly greater in the late-onset group than the early-onset group. Conclusion Patients should be monitored carefully for bile leakage for several weeks after hepatic resection, because late-onset bile leakage can cause serious complications. Intra-abdominal infection should also be treated as soon as possible, because it may induce refractory bile leakage with serious complications. Postoperative bile leakage can be a serious complication after hepatic resection. Few studies have analyzed patients according to the time of onset of bile leakage. We analyzed differences between patients with early- and late-onset bile leakage after hepatic resection and assessed clinical characteristics and outcomes in patients with late-onset leakage. Between 2008 and 2010, 1,009 patients underwent hepatic resection at 4 participating university hospitals and 2 community hospitals. Fifty-two patients (5.1%) with postoperative bile leakage were divided into an early-onset group (<2 weeks after surgery, n = 34) and a late-onset group (≥2 weeks after surgery, n = 18). Patient characteristics and outcomes were collected prospectively and analyzed retrospectively. The proportion of patients who underwent intra-abdominal placement of a drainage catheter was significantly less in the late-onset group than the early-onset group. All 18 patients in the late-onset group developed intra-abdominal infection, and 2 died of sepsis. The proportion of patients who underwent invasive treatment (abdominal paracentesis, endoscopic biliary drainage, or second hepatic resection) was significantly greater in the late-onset group than in the early-onset group. The time to resolution of bile leakage was significantly greater in the late-onset group than the early-onset group. Patients should be monitored carefully for bile leakage for several weeks after hepatic resection, because late-onset bile leakage can cause serious complications. Intra-abdominal infection should also be treated as soon as possible, because it may induce refractory bile leakage with serious complications.

Stress evokes lipolytic release of free fatty acid (FFA) and low-grade inflammation in visceral adipose tissue, mediated by increased adipokine secretion, and contributes to glucose metabolism disorder and prothrombotic state. We tested the hypothesis that alogliptin, a dipeptidyl peptidase-4 inhibitor, can ameliorate the biological effects of chronic stress in mice. C57BL/6J mice were subjected to 2-week intermittent restraint stress and orally treated with vehicle or alogliptin (dose: 15 or 45 mg/kg/day). Plasma levels of lipids, proinflammatory cytokines (monocyte chemoattractant protein-1, tumor necrosis factor-α, and interleukin-6), and 8-hydroxydeoxyguanosine were measured with enzyme-linked immunosorbent assay. Monocyte/macrophage accumulation in inguinal white adipose tissue (WAT) was examined by CD11b-positive cell count and mRNA expression of CD68 and F4/80 was examined by immunohistochemistry and RT-PCR, respectively. The mRNA levels of the above-mentioned proinflammatory cytokines, NADPH oxidase 4, adiponectin, and coagulation factors (plasminogen activation inhibitor-1 and tissue factor) in WAT were also assessed with RT-PCR. Glucose metabolism was assessed by glucose and insulin tolerance tests, plasma levels of DPP-4 activity, glucagon-like peptide-1, expression of DPP-4, insulin receptor substrate-1 and glucose transporter 4 in WAT and skeletal muscle. Alogliptin administration suppressed stress-induced FFA release, oxidative stress, adipose tissue inflammation, DPP-4 activation, and prothrombotic state in a dose-dependent manner, and improved insulin sensitivity in stressed mice. The results indicate that alogliptin improves stress-induced prothrombotic state and insulin resistance; suggesting that alogliptin could have beneficial therapeutic effects against cardiovascular complications in diabetic patients under stress.

Background: Laparoscopic hepatic resection (LHR) has been developed as a novel minimally invasive surgery. However, despite improvements in equipment and procedures, intraoperative hemorrhage remains an issue that requires great precaution. To reduce the amount of intraoperative blood loss, we perform the Pringle maneuver, aimed at occluding the inflow of blood into the liver during LHR. This article describes our experience performing LHR using the Pringle maneuver, including postoperative results, and discusses the safety and effectiveness of the Pringle maneuver. Methods: Data from 83 patients who underwent laparoscopic partial right hepatic resection with or without the Pringle maneuver were retrospectively analyzed with respect to surgical outcomes, safety, and utility. Results: In LHR, the amount of bleeding was significantly lower in cases that included the Pringle maneuver (P = .0314). However, there were no differences in the duration of surgery, surgical margin, rate of curative resections, and incidence of postoperative complications. Laboratory data collected after surgery showed no significant difference between the two groups regardless of whether blood flow was occluded or not. Conclusions: The Pringle maneuver may be effective in reducing the amount of intraoperative blood loss during laparoscopic partial right hepatic resection, although the difference is not clinically significant. Rather, the reduction in bleeding can reduce the stress experienced by the operator while keeping the transection stump of the liver dry. Particularly, the extracorporeal Pringle maneuver using cotton tape is simple and convenient and can be carried out within a short amount of time.

To determine the immune processes involved in chronic liver allograft rejection (CR) we examined in situ cytokine production in tissue from 15 patients with both clinical and histopathological diagnoses of CR. Total RNA was isolated from liver samples, reverse-transcribed and analyzed by RT-PCR for the production of proinflammatory cytokines and immunoregulatory mediators. Transcripts for the Th1-like cytokines IL-2 and IFN-gamma were detected in 53.3% and 46.7% of CR grafts, while they were detected in only 16% and 0% of stable grafts, respectively. The cytotoxic T cell mediator granzyme B was expressed in the majority of liver grafts undergoing CR, but was expressed only in a minority of stable grafts (80% vs. 16%, P < 0.05). The T cell product IL-5 was also significantly upregulated in CR as compared with stable livers (80% vs. 16%, P < 0.01). Other Th2 cytokines--IL-4 and IL-10--and macrophage products--IL-1 beta, IL-6, IL-8, TGF-beta, and TNF-alpha--were not substantially upregulated in CR grafts as compared with stable grafts. PDGF-beta transcripts were detected in the majority of the CR grafts, but were not detected in stable liver grafts (73% vs. 0, P < 0.05). By immunohistochemical staining, we observed that CD3+CD4+, and CD3+CD4- T cells were detected in CR grafts along with CD20+ B cells and CD68+ macrophages. There was, however, a predominant infiltration of CD3+CD4+ lymphocytes. Taken together, these data suggest that infiltrating cells produce proinflammatory and immunoregulatory cytokines that have a role in mediating graft damage in CR.

Although some previous studies have indicated the possibility of immunosuppression withdrawal in clinical liver transplantation, the mechanism of graft acceptance is not clear. The aim of this study is to elucidate the alloreactivity against the donor and intragraft cytokine profiles in living donor liver transplant (LDLT) recipients with graft acceptance. In October 1999, we had 23 patients who survived without immunosuppression after LDLT with a median drug-free period of 25 months (range: 3-69 months). They consisted of six patients who were electively weaned by an elective weaning protocol and 17 either forcibly or accidentally weaned patients due to various causes but mainly due to infection. We evaluated the alloreactivity against the donor in these patients by a mixed lymphocyte reaction and intragraft cytokine profiles by real-time reverse transcriptase-polymerase chain reaction. The development of donor-specific hyporeactivity was observed in the patients with graft acceptance. The cytokine pattern in the supernatant of the culture medium revealed a down regulation of T helper (Th) 1 cytokine INF gamma against the donor while no significant difference was seen in Th2 cytokine IL-10. Regarding the intragraft cytokine profiles, we could find no amplification of Thl cytokines (IL-2, INF y) and IL-4 while some of the patients revealed a gene expression of IL-10 with no significant difference from that of the normal, untransplanted liver specimen. In addition, no difference was observed in any other cytokines (IL-1beta, IL-8, IL-15, TNFalpha) compared with those of the normal controls. We propose that the down regulation of Th1 cytokine is one possible mechanism of graft acceptance in LDLT recipients.

The specific and efficient activation of mitogen-activated protein kinase (MAPK) signaling modules is mediated, at least in part, by scaffold proteins. c-Jun NH(2)-terminal kinase (JNK)-associated leucine zipper protein (JLP) was identified as a scaffold protein for JNK and p38 MAPK signaling modules. JLP is expressed nearly ubiquitously and is involved in intracellular signaling pathways, such as the G(alpha13) and Cdo-mediated pathway, in vitro. To date, however, JLP expression has not been analyzed in detail, nor are its physiological functions well understood. Here we investigated the expression of JLP in the mouse testis during development. Of the tissues examined, JLP was strongest in the testis, with the most intense staining in the elongated spermatids. Since the anti-JLP antibody used in this study can recognize both JLP and sperm-associated antigen 9 (SPAG9), a splice variant of JLP that has been studied extensively in primates, we also examined its expression in macaque testis samples. Our results indicated that in mouse and primate testis, the isoform expressed at the highest level was JLP, not SPAG9. We also investigated the function of JLP by disrupting the Jlp gene in mice, and found that the male homozygotes were subfertile. Taken together, these observations may suggest that JLP plays an important role in testis during development, especially in the production of functionally normal spermatozoa.

Arylamidase [EC 3.4.11.2] was isolated from monkey brain extract and purified about 2100-fold in approximately 11% yield by a six-step procedure comprising extraction from monkey brain homogenate, ammonium sulfate fractionation, first hydroxylapatite chromatography, DEAEcellulose chromatography, Sephadex G-200 gel filtration and second hydroxylapatite Chromatography. The enzyme showed a single band on polyacrylamide disc electrophoresis and consisted of a single polypeptide chain, as judged by disc electrophoresis in the presence of sodium dodecyl sulfate. The enzyme was strongly inhibited by PCMB, TPCK, and puromycin. Puromycin competitively inhibited the enzyme and the K1 value was about 5 × 10−7 M. Treatment with EDTA resulted in a loss of enzyme activity. The enzyme activity was restored by addition of Zn2+, Co2+, or Mn2+. Among various amino acid β-naphthylamides, L-alanine β-naphthylamide was most rapidly hydrolyzed and N-carbobenzoxyl-L-leucine β-naphthylamide was not hydrolyzed by this enzyme preparation. The molecular weight of the enzyme was 92,000 as determined by gel filtration on Sephadex G-200.

Abstract Introduction Heterotopic pancreatic cancer in the duodenum is a very rare disease. Only twelve cases have been reported worldwide to date. We report a rare case of malignant transformation of heterotopic pancreas (Heinrich type III) in the duodenum with long-term survival of the patient, and review the 12 cases in the literature. Case presentation A 75-year-old Japanese man was admitted to our hospital complaining of nausea and vomiting. Endoscopy and upper gastrointestinal contrast study showed marked duodenal stenosis. A pylorus-preserving pancreaticoduodenectomy was performed. Histopathological examination of the surgically resected specimen showed malignant transformation of heterotopic pancreas (Heinrich type III) in the duodenum. The postoperative course was uneventful, and the patient was discharged from the hospital on postoperative day 30. He is well and shows no signs of recurrence at the time of writing, six years after the surgery. Conclusion Adenocarcinoma arising within the heterotopic pancreas appears to be rare. It is difficult to obtain a correct diagnosis preoperatively. The management of heterotopic pancreas depends on the presence or absence of symptoms. If the patient is asymptomatic or benign, conservative treatment with regular follow-up is recommended. When the patient is symptomatic or there is a suspicion of malignancy, surgical management with intra-operative frozen section diagnosis is indicated.

In gastric cancer, various methods of gastric resection and reconstruction have been devised according to the location of the primary tumor and the depth of invasion. The functional outcomes of patients treated by laparoscopy-assisted or totally laparoscopic distal gastrectomy were compared with respect to the approach, size of the remnant stomach, and type of reconstruction.Patients who required distal gastrectomy to treat early-stage cancer between May 2000 and December 2008 were treated by one of the four following procedures: Billroth Type I (B-1) reconstruction for 1/2 remnant stomach (1/2B1ML) or B-1 for 1/3 remnant stomach (1/3B1ML), through a mini-laparotomy following laparoscopy-assisted surgery; intra-corporeal B-1 for 1/2 remnant stomach (1/2 B1IC); or intra-corporeal Roux-en-Y for 1/3 remnant stomach (1/3RYIC). The primary outcome measure was digestive function, assessed by body weight, food intake, and degree of abdominal symptoms. The secondary outcome was morbidity.The 1/2B1ML (n = 27) and 1/2B1IC (n = 56) groups were significantly superior to the 1/3 resection groups in terms of the preservation of body weight. The 1/3B1ML (n = 29) and 1/3RYIC (n = 64) groups were associated with significantly decreased food intake compared with the 1/2B1ML group. Endoscopy revealed a greater incidence of esophagitis and gastritis among the 1/3B1ML patients compared with the 1/3RYIC patients. There were no operative deaths, and no differences in morbidity between the groups.Patients with early-stage cancer actually benefit from 1/2 gastrectomy rather than the typical 2/3 gastrectomy. B-1 reconstruction is appropriate for patients with large gastric remnants, and intra-corporeal reconstruction in experienced hands is associated with no apparent disadvantages, while offering a favorable cosmetic result.

Abstract Background and Aim The evidence for main pancreatic duct intraductal papillary mucinous neoplasms ( MPD ‐ IPMN ) malignancy is based predominantly on investigation of resected cases, and the natural history is still unclear. The aim of the present study is to investigate the natural history of MPD ‐ IPMN and examine potential predictors of disease progression in MPD ‐ IPMN patients who conformed to “high‐risk stigmata” criteria. Methods This study included consecutive 20 follow‐up patients and 19 surgical patients with “high‐risk stigmata” MPD ‐ IPMN , in whom the diameter of the MPD was &gt; 10 mm, branch duct was &lt; 5 mm, and who underwent clinical follow up for ≥ 2 years. Results Among surgical patients, mural nodules and MPD diameter of invasive patients were significantly different compared with non‐invasive patients. On the other hand, among follow‐up patients, univariate analysis revealed the following potential predictors for disease progression: diameter of MPD ≥ 15 mm (hazard ratio [ HR ], 20.9; 95% confidence interval [ CI ], 2.59–173.4; P &lt; 0.01); and diffuse lesions of MPD ‐ IPMN ( HR , 4.46; 95% CI , 1.10–18.0; P = 0.04). On the other hand, multivariate analysis identified only diameter of MPD ≥ 15 mm ( HR , 19.2; 95% CI , 1.87–198.5; P = 0.01) as a potential predictor of disease progression. Conclusion If MPD ‐ IPMN patients have other severe complications or reasons for not undergoing surgical treatment, MPD diameter &lt; 15 mm, negative cytology, and no mural nodules, conservative clinical follow up for several years may be an option.

We report a very rare case of acute abdomen caused by torsion of an accessory spleen that was preoperatively diagnosed and cured by single-port surgery. A 31-year-old woman was admitted to our hospital with severe left abdominal pain. Physical examination revealed a left upper quadrant abdominal tenderness with voluntary guarding. Ultrasound demonstrated a well-defined round mass isoechoic to the spleen, measuring 3.0 cm in diameter in the left upper quadrant adjacent to the spleen. A contrast-enhanced CT scan showed a normally enhanced spleen and a 3.0 × 3.0, hypodense, non-enhancing mass anterior to the spleen with a twisted funicular structure. Torsion of an accessory spleen was suspected, and emergency single-port surgery was performed. During surgery, a rounded violet mass measuring 3.0 cm in diameter, suggestive of an accessory spleen, with a 1800° torsion around a long vascular pedicle along the left side of the greater omentum was discovered. The mass was removed and post-operative recovery was uneventful. A review of the literature revealed 26 cases (including ours) of torsion of an accessory spleen in English. Even with the recent advances in radiologic imaging modalities, making a preoperative diagnosis of this is difficult and most cases are diagnosed during laparotomy. This is the first report preoperatively diagnosed and cured by single-port surgery. We decided to start the operation by using a single port, not only for cosmetic reasons for this young female patient, but also for final confirmation of our diagnosis. We believe that single-port laparoscopy is valuable as a diagnostic tool as long as safety is assured for patients with acute abdomen. Although torsion of an accessory spleen is extremely rare, it should be considered in the differential diagnosis of acute abdomen in children and young adults.

The effect of the chymase inhibitor TY-51469 on the development and progression of non-alcoholic steatohepatitis (NASH) was evaluated in rats fed a high-fat and high-cholesterol (HFC) diet. To evaluate the preventive effect of TY-51469 on the development of NASH, stroke-prone spontaneously hypertensive rat 5 (SHRSP5)/Dmcr rats were fed either a normal or HFC diet for 8 weeks, and concurrently administered either placebo or TY-51469 (1 mg/kg per day). To evaluate the effect of TY-51469 on the survival rate, TY-51469 was administered either concurrently with HFC diet (pretreated group) or 8 weeks after HFC diet at which point NASH had developed (posttreated group). Eight weeks after HFC diet, significant increases of steatosis, fibrosis and chymase-positive cells were observed in liver from the placebo-treated rats. Significant increases of myeloperoxidase, transforming growth factor-β, matrix metalloproteinase-9, and collagen I mRNA levels were also observed. However, all parameters were significantly attenuated in the TY-51469-treated group. A survival rate of the placebo-treated group fed the HFC diet was 0% at 14 weeks. In comparison, the rates of TY-51469-pretreated and TY-51469-posttreated groups were 100% and 50% at 14 weeks, respectively. Chymase inhibitor may be applicable to preventing the development and progression of NASH.

We examined the expression of full-length TrkB (TrkBTK+) and truncated TrkB (TrkBTK−) in the central nervous system (CNS) of the macaque monkey (Macaca fascicularis) using a western blot analysis. At the adult stage, the levels of TrkBTK+ in cerebral cortices were higher than those in other structures of CNS and the expressions of TrkBTK+ in the association cortices (except area PE) were relatively lower than those in the primary cortices. In contrast, TrkBTK− in the hippocampus and the cerebellum was significantly higher than in other structures. In various developing cerebral cortices, TrkBTK+ was detected at the same levels from embryonic day 120 (E120) to the adult period. In contrast, the expression of TrkBTK− increased remarkably after the newborn stage (NB), reached the maximum level at postnatal day 60 (P60) and maintained the same level into adulthood. The peaks of TrkBTK− in the association cortices were more delayed than in the primary cortices. The expression of TrkBTK− occurred at a time that correlates with the elimination of axons and the down-regulation of neuropeptides. The present study suggests that TrkBTK− plays an important role in the axonal remodelling and that it may act as a negative effector of TrkBTK+ in the primate CNS, reducing responsiveness to BDNF and/or NT-4/5.

Background & Aims: More than 2000 adult-to-adult living donor liver transplantations (LDLT) have been performed in the United States, yet the potential benefit to liver transplant candidates of undergoing LDLT compared with waiting for deceased donor liver transplantation (DDLT) is unknown. The aim of this study was to determine whether there is a survival benefit of adult LDLT. Methods: Adults with chronic liver disease who had a potential living donor evaluated from January 1998 to February 2003 at 9 university-based hospitals were analyzed. Starting at the time of a potential donor’s evaluation, we compared mortality after LDLT to mortality among those who remained on the waiting list or received DDLT. Median follow-up was 4.4 years. Comparisons were made by hazard ratios (HR) adjusted for LDLT candidate characteristics at the time of donor evaluation. Results: Among 807 potential living donor recipients, 389 underwent LDLT, 249 underwent DDLT, 99 died without transplantation, and 70 were awaiting transplantation at last follow-up. Receipt of LDLT was associated with an adjusted mortality HR of 0.56 (95% confidence interval [CI]: 0.42–0.74; P < .001) relative to candidates who did not undergo LDLT. As centers gained greater experience (>20 LDLT), LDLT benefit was magnified, with a mortality HR of 0.35 (95% CI: 0.23–0.53; P < .001). Conclusions: Adult LDLT was associated with lower mortality than the alternative of waiting for DDLT. This reduction in mortality was magnified as centers gained experience with LDLT. This reduction in transplant candidate mortality must be balanced against the risks undertaken by the living donors themselves.

BDNF and its specific receptor TrkB are concerned with synaptic plasticity as well as maintenance of the nervous system. TrkB has three subtypes: full-length TrkB (TK+), which has a tyrosine kinase containing intracellular domain, and two truncated TrkBs (TK−; T1 and T2), which lack tyrosine kinases. To understand the molecular interaction among these subtypes, we investigated the expression and distribution of BDNF, TK+, and T1 in the adult monkey cerebellum by single and double immunohistochemistry and Western blot analysis. We observed by single immunohistochemistry that BDNF, TK+, and T1 are distributed in almost all the somata and dendrites of Purkinje and granule cells. In the double-stained sections, three kinds of regions were observed: TK+ >T1; TK+ =T1; TK+ <T1. Moreover, three types of TrkB dimers (TK+/TK+ homodimer, TK+/TK− heterodimer, and TK−/TK− homodimer) were induced by stimulating with exogenous BDNF. These observations suggest that the functions of BDNF may be modified by interaction among subtypes of TrkB in each region of the Purkinje cells.

Rat pleurisy induced by carrageenin or phorbol myristate acetate (PMA) was utilized for examination of the plasma exudation process of inflammation. Chemical mediators responsible for induction of vascular permeability increase were examined. In carrageenin-induced pleurisy, kinin and PGI&lt;sub&gt;2&lt;/sub&gt; were demonstrated as the main mediators, and in PMA pleurisy histamine, PGI&lt;sub&gt;2&lt;/sub&gt; and platelet-activating factor were the main mediators. These results indicate that different stimuli may activate different enzymatic processes to produce different mediators, but they may result in similar inflammatory reactions by action of these multiple mediators simultaneously released.

Aim: Mast cells may be involved in the pathogenesis of nonalcoholic steatohepatitis (NASH). The mast cell protease chymase contributes to the formation of angiotensin II and matrix metalloproteinase (MMP)‐9, both of which are intimately involved in liver fibrosis. Therefore, we hypothesized that chymase plays an important role in the development of NASH. Methods: Hamsters were fed a methionine‐ and choline‐deficient (MCD) diet for 8 weeks. These animals were divided into two groups and received either TY‐51469 (1 mg/kg per day) or placebo. A third group was fed a normal diet as a control. Results: Total plasma bilirubin, triglycerides, and hyaluronic acid levels were significantly higher in the MCD diet‐fed hamsters than in the normal diet‐fed hamsters, but the levels were significantly lower in chymase inhibitor‐treated MCD diet‐fed hamsters than in placebo‐treated MCD diet‐fed hamsters. Using histological analysis, marked steatosis and fibrosis were observed in MCD diet‐fed hamsters, but these changes were significantly attenuated by treatment with the chymase inhibitor. Increases in mast cells and chymase‐positive cells were observed in the liver after the MCD diet, but the increases disappeared in the chymase inhibitor‐treated group. The significant increase observed in chymase activity in liver tissue extract from the MCD diet‐fed group was also reduced by treatment with the chymase inhibitor. Chymase inhibition significantly reduced not only angiotensin II expression but also matrix metallopeptidase 9 activity in MCD diet‐fed hamsters. Conclusion: These findings demonstrate that the mast cell protease chymase may play a crucial role in the development of NASH in hamsters.

Aim: Chymase converts angiotensin I to angiotensin II, which may promote the development of liver fibrosis. In this study, whether a chymase inhibitor TY‐51469 attenuated tetrachloride (CCl 4 )‐induced liver fibrosis was examined. Methods: Liver fibrosis was induced by the s.c. injection of 1 mL/kg of CCl 4 twice weekly for 8 weeks, and each hamster was given TY‐51469 (1 mg/kg per day) or placebo. Untreated hamsters were used as a control group. Results: Significant increases of serum alanine aminotransferase, total bilirubin and hyaluronic acid levels were observed in the placebo‐treated group compared with the control group, but these levels were significantly attenuated in the TY‐51469‐treated group. Liver chymase activity was significantly higher in the placebo‐treated group than in the control group, whereas the activity in the TY51469‐treated group was not. Total angiotensin II‐forming activity in the liver was also significantly higher in the placebo‐treatedgroup than in the control group or the TY‐51469‐treated group. The ratio of the fibrotic area to the total area in the liver was significantly higher in the placebo‐treated group than in the control group, but the ratio was significantly lower in the TY‐51469‐treated group than in the placebo‐treated group. A significant decrease in the number of α‐smooth muscle actin (SMA)‐positive cells was seen in the TY‐51469‐treated group compared to the placebo‐treated group. Conclusion: Significant correlations between the number of chymase‐positive cells and the degree of fibrosis and between the numbers of chymase‐positive cells and α‐SMA‐positive cells were observed. Thus, chymase inhibition may be a useful strategy for preventing liver fibrosis.

Abstract Background Previously, we demonstrated decreased expression of somatostatin mRNA in aged macaque brain, particularly in the prefrontal cortex. To investigate whether or not this age‐dependent decrease in mRNA is related to morphological changes, we analyzed somatostatin cells in the cerebra of aged Japanese macaques and compared them with those in rats and tree shrews, the latter of which are closely related to primates. Methods Brains of aged macaques, tree shrews, and rats were investigated by immunohistochemistry with special emphasis on somatostatin. Results We observed degenerating somatostatin‐immunoreactive cells in the cortices of aged macaques and tree shrews. Somatostatin‐immunoreactive senile plaque‐like structures were found in areas 6 and 8 and in the nucleus accumbens of macaques, as well as in the nucleus accumbens and the cortex of aged tree shrews, where amyloid accumulations were observed. Conclusions Somatostatin degenerations may be related to amyloid accumulations and may play roles in impairments of cognitive functions during aging.

Radiofrequency catheter ablation (RFCA) is increasingly performed for the treatment of atrial fibrillation (AF), but it is problematic because the use of anti-arrhythmic agents is largely restricted in patients undergoing hemodialysis (HD) therapy. However, little is known about the long-term clinical outcomes of AF after RFCA in HD patients.Between 2002 and 2008, 16 HD patients (age: 63.8 ± 7.4 years, 75.0% men) underwent RFCA for AF at the Toyota Kosei Hospital. We investigated the long-term results and mortality of RFCA for AF in HD patients and compared them with those of 111 non-HD patients (age: 58.6 ± 10.0 years, 78.3% male) who received the same procedures.During the follow-up (64.3 ± 25.4 months in HD patients, 70.5 ± 20.2 months in non-HD patients) after the initial RFCA procedure, sinus rhythm was restored in 4 HD patients (25%) and in 45 non-HD patients (40.5%). Multiple procedures were performed in 12 HD patients and in 57 non-HD patients. After the final procedure, 13 HD patients (81.3%) and 92 non-HD patients (82.9%) were free of atrial arrhythmia and symptoms. Of importance, Kaplan-Meier analysis did not demonstrate any significant differences in the atrial arrhythmia-free rate after the last procedure between HD patients and the control group matched after propensity-score analysis despite higher all-cause mortality in HD patients than in non-HD patients.During 5-years of follow-up, the use of multiple RFCA procedures for AF in patients undergoing HD was favorable, whereas the use of a single procedure was disappointing. Multiple RFCA procedures can be an efficient approach to the treatment of AF in HD patients.

Purpose The purpose of this retrospective study was to evaluate the utility of the new intraoperative bile leakage test as a preventive measure of postoperative bile leakage. Method 737 patients were retrospectively analyzed with respect to the management of intra- and post-operative bile leakage. Results Nine (8.3%) of 109 patients evaluated using conventional white light fluorescent imaging were recognized as having intra-operative bile leakage. However, performance of 5-aminolevulinic acid (5-ALA)-mediated PDD detected bile leakage intraoperatively not only in these 9 patients, but also in an additional 6 patients, such that ‘red fluorescence’ at the cut surface of the liver, was visualized in a total of 15 patients. The postoperative courses of most patients were uneventful, and postoperative bile leakages occurred in only one (0.9%) patient. Conclusion 5-ALA fluorescence imaging may be needed to prevent postoperative bile leakage in patients at high risk for this surgical complication after hepatic resection.

Bevacizumab plus paclitaxel therapy for recurrent breast cancer did not prolong overall survival(OS)in clinical trials, but it was efficacious for treating life-threatening HER2-negative recurrent breast cancer. This regimen is often used in daily clinical practice by breast surgeons. However, bevacizumab therapy results in unique adverse events, of which proteinuria and hypertension are relatively frequent. Moreover, the symptoms often improve on reducing the dose or discontinuing the drug. In this case, bevacizumab administration caused delayed wound healing, making subsequent anticancer treatment difficult, and consequently we could not prolong the patient's life.

We have adopted the use of an oral tacrolimus induction protocol in pediatric liver transplantation since the commercial release of tacrolimus in 1994. In this study we analyzed the efficacy of oral tacrolimus induction therapy in 17 consecutive transplants (15 patients) performed between 6/94 and 2/95 and 4 additional patients who were retransplanted between 11/93-5/94 and received compassionate oral tacrolimus induction. Sixteen transplants were treated with oral tacrolimus induction only; 5 transplants, oral tacrolimus + ATGAM/OKT3 induction. The protocol consisted of 0.2 mg/kg of tacrolimus orally on the first postoperative day with a corticosteroid taper. Oral tacrolimus was started at day 1-8 in the 5 patients receiving ATGAM/OKT3 induction. Dosages were adjusted over time to maintain a whole-blood trough level of 12-15 ng/ml at 0-1 month, 10-12 ng/ml at 1-3 months, and 5-10 ng/ml after 3 months. The incidence of acute rejection was 50% (8/16) in children on oral tacrolimus induction alone and 80% (4/5) in the tacrolimus + ATGAM/OKT3 group. Epstein-Barr virus infection occurred in 6 of 19 children (32%), with no child developing lymphoproliferative disorder. No adverse effect on renal function was noted. Serum fasting glucose was stable over time while a trend was noted in decreasing serum cholesterol levels at 6 months. Antihypertensive medication was required in 4 of 19 children (21%) posttransplantation. Corticosteroids were withdrawn in 11% (2/19) of patients. Actuarial 1-year patient and graft survivals were 95% and 86%, respectively. The use of oral tacrolimus induction therapy was associated with excellent survival and a low incidence of complications.

We performed nonradioactive in situ hybridization histochemistry (ISH) in the lateral geniculate nucleus (LGN) of the macaque monkey to investigate the distribution of mRNA for two growth-associated proteins, GAP-43 and SCG10. GAP-43 and SCG10 mRNAs were coexpressed in most neurons of both magnocellular layers (layers I and II) and parvocellular layers (layers III-VI). Double-labeling using nonradioactive ISH and immunofluorescence revealed that both GAP-43 and SCG10 mRNAs were coexpressed with the alpha-subunit of type II calcium/calmodulin-dependent protein kinase, indicating that both mRNAs are expressed also in koniocellular neurons in the LGN. We also showed that GABA-immunoreactive neurons in the LGN did not contain GAP-43 and SCG10 mRNAs, indicating that neither GAP-43 nor SCG10 mRNAs were expressed in inhibitory interneurons in the LGN. GABA-immunoreactive neurons in the perigeniculate nucleus, however, contained both GAP-43 and SCG10 mRNAs, indicating that both mRNAs were expressed in inhibitory neurons in the perigeniculate nucleus, which project to relay neurons in the LGN. Furthermore, to determine whether the expression of GAP-43 and SCG10 mRNAs is regulated by visual input, we performed nonradioactive ISH in the LGN and the primary visual area of monkeys deprived of monocular visual input by intraocular injections of tetrodotoxin. Both mRNAs were downregulated in the LGN after monocular deprivation for 5 d or longer. From these results, we conclude that both GAP-43 and SCG10 mRNAs are expressed in the excitatory relay neurons of the monkey LGN in an activity-dependent manner.

Non-invasive methods for monitoring reproductive status of chimpanzee based on the measurement of urinary steroids and gonadotropins were examined. A typical pre-ovulatory urinary estrone conjugate (E1C) surge and post-ovulatory increase in pregnandiol glucuronide (PdG) were seen during the menstrual cycle. Urinary follicle stimulating hormone (FSH) showed two peaks over the infertile menstrual cycle. The earliest changes indicating pregnancy were a coincident rise in E1C and chorionic gonadotropin (CG) levels and a concomitant fall in FSH levels. Urinary PdG levels showed a prolonged rise. Urinary E1C in the pregnant chimpanzee was higher than during the menstrual cycle and increased with advancing gestation, with maximum levels occurring near term. In the case of stillbirth, E1C and CG levels from mid- through late-pregnancy were low and the prepartum progressive increase in E1C was not shown. The data presented here are of great practical value in captive breeding management of chimpanzees.

An enzyme immunoassay for nerve growth factor was developed to determine the regional distribution and ontogenic change in the macaque (Macaca fascicularis) CNS. The standard curve of mouse nerve growth factor paralleled the dilution curves of extracts from the primate CNS at the adult and pre-natal stages. Furthermore, the nerve growth factor immunoreactive material comigrated with mouse nerve growth factor by means of carboxy methyl cellulose chromatography. These findings suggest that the immunoreactive material extracted from the primate CNS is mouse nerve growth factor-like molecules. At the adult stage, the highest level of nerve growth factor was in the hippocampus, with relatively high levels also in the hypothalamus, the cerebral cortex, the amygdala, the basal nucleus of Meynert, the septal nucleus, the cerebellum and the caudate nucleus. No detectable amounts were observed in the spinal cord, the substantia nigra or the dentate nucleus. In addition to the CNS, the pituitary gland contained about four times the level found in the hippocampus. At embryonic day 120, a high level of nerve growth factor already existed in the occipital cortex (80% of the level at the adult stage) and in the hippocampus (70% of the level at the adult stage). Between embryonic day 120 and the newborn stage in the occipital cortex and between embryonic day 120 and postnatal day 60 in the hippocampus, nerve growth factor levels increased about 1.7-fold, and after that, they gradually decreased until the adult stage was reached. In contrast, in the cerebellum, the level was quite high during the pre-natal period and declined to one-third at postnatal day 60. The developmental changes in nerve growth factor and choline acetyltransferase activity in the hippocampus were well correlated (r = 0.963) between embryonic day 120 and postnatal day 60. Our studies reveal that nerve growth factor is present in the primate CNS. The high level of nerve growth factor during embyronic stages and the good correlation with choline acetyltransferase activity suggest a physiological role for nerve growth factor in the development of the primate CNS.

Much attention has focused on tachykinin receptors as therapeutic targets for neuropsychiatric disorders, although their expressional distributions in the primate central nervous system (CNS) remain unclear. We cloned the genes encoding the NK‐1 and NK‐3 tachykinin receptors (referred to as rmNK‐1 and rmNK‐3) from the rhesus monkey ( Macaca mulatta) brain and examined their pharmacological profiles and regional distributions in the CNS. The deduced rmNK‐1 amino‐acid sequence differed by only two amino acids from the human NK‐1 (hNK‐1). The deduced rmNK‐3 amino‐acid sequence was two amino acids shorter than human NK‐3 (hNK‐3), with a seven‐amino‐acid difference in sequence. Ligand binding studies revealed that the affinity of rmNK‐1 to substance P (SP) was comparable to that of hNK‐1 in cell lines that expressed individual receptors stably. Nonpeptide antagonists had similar effects on the binding of rmNK‐1 and hNK‐1. Affinity of rmNK‐3 for NKB was stronger than for SP and the IC 50 value was comparable with that of hNK‐3. Ca 2+ imaging showed that activations of both rmNK‐1 and rmNK‐3 by specific ligands, SP and senktide, induced increased intracellular Ca 2+ in cell lines that stably expressed individual primate tachykinin receptors. The amounts of rmNK‐1 and rmNK‐3 mRNAs were quantitatively determined in the monkey CNS. The expression of rmNK‐1 was observed in all of the cortical and subcortical regions, including the hippocampus and the amygdala. The putamen contained the most NK‐1 mRNA in the brain, with less rmNK‐3 mRNA found in the cortex compared to rmNK‐1 mRNA. In the monkey hippocampus and amygdala, rmNK‐1 mRNA was present at markedly higher concentrations than rmNK‐3 mRNA. The present results provide an insight into the distinct physiological nature and significance of the NK‐1 and NK‐3 tachykinin systems in the primate CNS. These findings are indispensable for establishing model systems in the search for a subtype‐specific tachykinin receptor agonist and antagonist for the treatment of neuropsychiatric disorders. British Journal of Pharmacology (2006) 147 , 316–323. doi: 10.1038/sj.bjp.0706561

A truncated TrkB receptor, T1, which is one of the receptors for brain-derived neurotrophic factor, has been shown to regulate the morphology of neurons and glial cells in primary cultures and/or slices overexpressing T1 in the recent past. However, in vivo localization of T1 at protein level remains unclear. In the present study, we examined the localization of T1 in the primary motor and prefrontal cortices of adult monkeys by using immunohistochemistry. In the primary motor cortex, T1 immunoreactivity was observed mainly in the pyramidal neurons of layers II-VI, especially Betz cells of layer V. The apical and basal dendrites and cell bodies of Betz cells were strongly stained. In addition, we found that the interneurons were also T1-immunopositive and that there were no T1-positive astrocytes. In the prefrontal cortex, we observed strong immunoreactivity of T1 in astrocytes as well as pyramidal neurons of layer V. The pyramidal neurons and interneurons in layers II/III were faintly immunoreactive for T1. Thus, these findings, together with the fact that T1 is involved in morphological control of neurons and glial cells, suggest that the prefrontal cortex might possess a different degree of morphological plasticity than the primary motor cortex in the adult monkey.

From the perspective of comparative morphology, the distribution of non-monoaminergic neurons in the common marmoset (Callithrix jacchus) was investigated using an immunohistochemical method with specific antibodies to tyrosine hydroxylase (TH) and aromatic-L-amino acid decarboxylase (AADC). TH-immunoreactive (IR) neurons (but not AADC-IR) neurons were observed in the olfactory tubercle, preoptic suprachiasmatic nucleus, periventricular hypothalamic nucleus, arcuate nucleus, paraventricular nucleus, periaqueductal gray matter, medial longitudinal fasciculus, substantia nigra, and nucleus solitaris. In contrast, AADC-IR (but not TH-IR), small, oval and spindle-shaped neurons were sparsely distributed in the following areas: the hypothalamus from the anterior nucleus to the lateral nucleus, the dorsomedial nucleus, the dorsomedial area of the medial mammillary nucleus and the arcuate nucleus; the midbrain, including the stria medullaris and substantia nigra; and the medulla oblongata, including the dorsal area of the nucleus solitaris and the medullary reticular nucleus. The distribution of AADC-IR neurons was not as extensive in the marmoset as it is in rats. However, these neurons were located in the marmoset, but not the rat substantia nigra. Furthermore, AADC-IR neurons that are present in the human striatum were absent in that of the marmoset. The present results indicate that the distribution of non-monoaminergic neurons in the brain of the common marmoset is unique and different from that in humans and rodents.

Aim: Angiotensin II may contribute to liver fibrogenesis. In addition to angiotensin‐converting enzyme (ACE), chymase, which is expressed by mast cells, is also known to be an angiotensin II‐forming enzyme. However, it is unclear which of these two angiotensin II‐forming enzymes plays a more important role in liver cirrhosis progression. In the present study, the role of angiotensin II‐forming enzymes in the progression of liver cirrhosis was clarified. Methods: A total of 77 patients (16 in F0 stage, 10 in F1 stage, 22 in F2 stage, 12 in F3 stage, and 17 in F4 stage) were classified according to the new Inuyama classification into a non‐cirrhosis (F0) group, an early cirrhosis (F1 + F2) group, and a chronic cirrhosis (F3 + F4) group. Results: Both chymase and total angiotensin II‐forming activities were significantly higher in chronic cirrhosis patients than in the other two groups. However, there was nodifference among the three groups in ACE activity. On immunohistology, the number of chymase‐ and angiotensin II‐positive cells was significantly higher in the chronic cirrhosis group than in the non‐cirrhosis and early cirrhosis groups. There were significant correlations between the number of chymase‐positive cells and the number of angiotensin II‐positive cells, between the number of chymase‐positive cells and the degree of fibrosis, and between the number of angiotensin II‐positive cells and the degree of fibrosis. Conclusion: These results suggest that chymase‐dependent angiotensin II formation may play an important role in hepatic fibrosis of patients with cirrhosis.

Abdominal drainage after liver resection is considered unnecessary: however, there still exist a number of cases where drain is effective to prevent serious infectious complications. We re-evaluated the necessity of drain placement after liver resection from the retrospective analysis of postoperative complications with special reference to the need for drain insertion of 140 patients undergoing hepatectomy without intraoperative abdominal drainage from 2007 through 2010. Three patients required drain reinsertion in the early postoperative period (before postoperative Day 7); all had undergone extended right hepatectomy for hepatocellular carcinoma with portal vein thrombus followed by postoperative liver failure. Risk factors for postoperative bile leakage included repeated hepatectomy, operative procedure with exposure of the major Glisson's sheath (i.e., central bisegmentectomy and anterior segmentectomy), and intraoperative bile leakage. However, because the onset of this complication was as late as postoperative Day 19.5, prophylactic drainage does not appear useful. Although not required routinely, prophylactic drainage might be useful in patients undergoing extended hepatectomy, a high-risk hepatectomy procedure exposing the major Glisson's sheath, those with positive intraoperative bile leakage, for hepatocellular carcinoma, and especially complicated with portal vein thrombus.

Changes in the expression of estrogen-related substances in monkeys' brains at the menopausal transition, when estrogen deficit starts to occur, have not yet been examined thoroughly. In the present study, we immunohistochemically investigated the expression levels of estrogen receptor beta (ERβ) and aromatase (local estrogen synthesizing enzyme) in the hippocampal formation of premenopausal, menopausal, and ovariectomized premenopausal monkeys. In all monkeys tested, ERβ immunoreactivity was observed in interneurons located in the subiculum and the Ammon's horn, and most of these ERβ-immunoreactive neurons coexpressed a GABAergic neuron marker, parvalbumin. In the menopausal monkeys who exhibited a decline in estrogen concentration, hippocampal ERβ was highly upregulated, while aromatase expression was not markedly changed. By contrast, aromatase in the ovariectomized monkeys was significantly upregulated, while ERβ expression was not changed. In the brains of ovariectomized and menopausal monkeys, depletion of ovary-derived estrogen brought about different reactions which may be attributed to the senescence of brain aging.

Because pancreatic cancer is a disease with a dismal prognosis due to the high rate of early recurrence even after curative surgery, selecting the most effective treatment in an individual requires preoperative assessment of the tumor characteristics, including the potential for early recurrence. The study cohort included 84 patients undergoing surgical resection of pancreatic cancer. Univariate and multivariate analyses were conducted to identify the risk factors for early recurrence within six months after curative resection. Early recurrence was associated with a platelet-lymphocyte ratio ≥0.23 (P = 0.04), carbohydrate antigen 19-9 (CA19-9) ≥200 (P = 0.01), and S-pancreas-1 antigen (Span-1) ≥37 (P = 0.0004) by univariate analysis. Multivariate analysis identified CA19-9 ≥200 and Span-1 ≥37 as independent risk factors for early recurrence. Patients with both risk factors had a significantly higher rate of lymph node metastasis than those with no or one risk factor. Span-1 ≥ 37 and CA19-9 ≥ 200 are independent risk factors for early recurrence in patients who underwent surgical resection, and the combination of Span-1 ≥37 and CA19-9 ≥200 is a useful indicator of lymph node metastasis.